Library-free methylation sequencing with bisulfite padlock ... · Nature Methods Library-free...

Nature Methods

Library-free methylation sequencing with bisulfite padlock

probes

Dinh Diep, Nongluk Plongthongkum, Athurva Gore, Ho-Lim Fung, Robert Shoemaker & Kun Zhang

Supplementary Figure 1 Schematic for the probe design software (ppDesigner)

Supplementary Figure 2 Schematic for the bisulfite sequencing data analysis pipeline

(bisReadMapper)

Supplementary Figure 3 Comparison of probe capture efficiencies between the DMR220K, LC4K

probe sets and the previously published CGI30K set

Supplementary Figure 4 Scatter plot of number of characterized CpG sites versus mappable

sequencing data for the DMR330K probe set

Supplementary Figure 5 Number CpG sites called per sample as a function of sequencing effort

Supplementary Figure 6 Captured CpG sites were tested for potential regulatory interactions with

genes by GREAT

Supplementary Figure 7 Accuracy of digital quantification of BSPP

Supplementary Figure 8 Comparison between BSPP and whole genome bisulfite sequencing (WGBS)

Supplementary Figure 9 Variation in amount of sequencing data obtained per sample in a multiplex

BSPP capture experiment

Supplementary Figure 10 Example padlock probes

Supplementary Figure 11 UCSC Genome Browser view showing an example of aberrant iPSC-specific

methylation after reprogramming of PGP1 fibroblasts into iPS cells

Supplementary Table 1 Comparison of bisulfite sequencing methods

Supplementary Table 2 Representative cost per sample

Supplementary Table 3 Primer sequences used

Supplementary Table 4 Sequences of multiplexed barcoded primers

Nature Methods: doi:10.1038/nmeth.1871

1

Supplementary Figure 1. Schematic for the probe design software (ppDesigner). The neural network model utilizes the target length, target GC content, binding arm melting temperature, binding arm length, local single-stranded folding energy of the target, and the dinucleotides present at the extension site and ligation site during probe capture. Example probes can be found in Supplementary Figure 10.


2

Supplementary Figure 2. Schematic for the bisulfite sequencing data analysis pipeline

(bisReadMapper).


3

Supplementary Figure 3. Comparison of probe capture efficiencies between the DMR220K, LC4K

probe sets and the previously published CGI30K set. The first three plots were generated from data

without subsetting or suppressor oligos to allow for a direct comparison of probe design.


4

Supplementary Figure 4. Scatter plot of number of characterized CpG sites versus mappable

sequencing data for the DMR330K probe set. Variability in sequencing quality of individual sequencing

runs is responsible for the different number of CpG sites characterized with similar sequencing effort.

0

100,000

200,000

300,000

400,000

500,000

600,000

700,000

0 0.5 1 1.5 2 2.5 3 3.5

Nu

mb

er

of

Cp

Gs

Ch

ara

cte

rize

d

Bases Mapped (Gbps)

Library-free PE 110bp reads

(n=16)

Library-free PE 100 bp reads

(n=46)


5

Supplementary Figure 5. Number CpG sites called per sample as a function of sequencing effort.

The horizontal dash line represents 4Gbps of sequences per library that we routinely generate.

0

200000

400000

600000

800000

1000000

1200000

0.1 1 10 100

Nu

mb

er

of

On

-Targ

et

Cp

Gs C

hara

cte

rized

Sequencing Effort (Gbps)

1x Sequencing Depth

5x Sequencing Depth

10x Sequencing Depth

Typical Sequencing Effort


6

Supplementary Figure 6. Captured CpG sites were tested for potential regulatory interactions with

genes by GREAT (http://great.stanford.edu). (A) Most CpG sites were interacting with 1-2 genes. (B)

Distance of CpG sites to the transcriptional start sites (TSS) of the predicted regulating genes.


http://great.stanford.edu/

7

Supplementary Figure 7. Accuracy of digital quantification by BSPP. (a,b) Within batch and between

batch comparison of the methylation levels obtained at 10x depth from multiple capture reactions of

the same sample (PGP1iPS). The Pearson’s correlation coefficient R for within one batch is 0.98

(N=405,508), and for between batches is 0.97 (N=117,186). (c,d,e) Within sample comparison of

methylation levels obtained from different probes capturing the same CpG site on different strands at

10x depth within one capture reaction. The Pearson’s correlation coefficient R was 0.96 (N=44,361),

0.96 (N=55,965), and 0.97 (N=29,884) for PGP1iPS, PGP1F, and H1 respectively.


8

Supplementary Figure 8. Comparison between BSPP and whole genome bisulfite sequencing

(WGBS). We compared two H1 ESC datasets, using sites with at least 10x read depth in each. The

Pearson’s correlation coefficient R was 0.9477 (N=135,300). (Note that the sequencing experiments

were performed on separate cultures of H1 from two different labs.)

N = 135,300 r = 0.9477


9

Supplementary Figure 9. Variation in amount of sequencing data obtained per sample in a

multiplexed BSPP capture experiment. 48 whole blood samples were captured and sequenced in one

batch using the library-free BSPP method. There is little variation between samples in the amount of

generated sequencing data.


10

Supplementary Figure 10. Example padlock probes ordered from (A) Agilent’s oligonucleotide

synthesis service and (B) LC Sciences’ oligonucleotide synthesis service.


11

Supplementary Figure 11. UCSC Genome Browser view showing an example of aberrant iPSC-

specific methylation after reprogramming of PGP1 fibroblasts into iPS cells. Circles represent a

location with measurable methylation state, with black indicating unmethylated and gold indicating

methylated. The Agilent 330K probe set identified a small intronic region containing aberrant

methylation in the iPS cells that are not present in either the fibroblast progenitors or a control hESC

line. The LC Sciences 4K probe set was designed to characterize the methylation state upstream and

downstream of this region. This focused assay revealed that the abnormal methylation also extended

into the exonic region of GRM7.


12

Supplementary Table 1. Comparison of bisulfite sequencing methods. The number of enzymatic

reactions, number of purifications, cost per sample, and mapping rates for first-generation padlock

probes, second-generation library-free padlock probes, reduced representation bisulfite sequencing

(RRBS), and whole genome bisulfite sequencing (WGBS) are shown.

Published BSPP

Library-free BSPP

RRBS WGBS

Enzymatic reactions 10 3 4 3

Purification 6 1 3 3

Size-selection 2 11 1 1

Sample preparation cost per sample

$71.151 $37.86

2 $28.15 $31.10

Mapping rate 44% 87% 27%3 N.D.

Genome coverage obtained at 10x depth

<0.1%

0.6%-1%

~1%3 76-96%

4

Sequencing (Gbps) 0.5 4.0 1.4 70.0

Sequencing cost per sample

5

$24.38 $195.00 $68.25 $3412.50

1Unlike other methods, in the library-free BSPP protocol size selection is typically performed on 48-96 pooled

libraries. 2 Includes the cost of ordering 400,000 synthesized probes from LC Sciences and reagents for preparing probes,

bisulfite conversion, capture, and sequencing library preparation. Estimates assume that 10,000 samples will be processed. 3Estimated from: Gu et. al., Nat Methods 2010; 7(2):133-136.

4 Adapted from: Beck et. al., Nat Biotechnol 2010;28:1026-1028.

5 Assumes sequencing using an Illumina HiSeq to generate 300 Gbps of sequencing data, with cost of $4920 for

a flowcell, $6815 for sequencing reagents, and $2890 for service fee. ($48.75 per Gbps)


13

Supplementary Table 2. Representative cost per sample for oligonucleotide synthesis, sequencing library construction, and Illumina sequencing.

Expected number of samples to be processed

Probe set sizes

4,000 40,000 400,000

10 $134.57 $872.04 $9,298.78

100 $35.57 $129.54 $1,131.28

1000 $25.67 $55.29 $314.53

10000 $24.68 $47.86 $232.86


14

Supplementary Table 3. Primer sequences used for padlock probe production, padlock capture, sequencing library construction, and Illumina sequencing.

Primer name Primer sequences Primers used with Agilent Probes pAP1V61U 5’-G*G*G*TCATATCGGTCACTGTU-3’ AP2V6 5’-/5Phos/CACGGGTAGTGTGTATCCTG-3’ RE-DpnII-V6 5’-GTGTATCCTGATC-3’

AmpF6.4Sol 5’-AATGATACGGCGACCACCGAGATCTACACCACTCTCAGATGTTATCGAGGTCCGAC-3’

AmpF6.3NH2 5’-/5AmMC6/CAGATGTTATCGAGGTCCGAC-3’ AmpR6.3NH2 5’-/5AmMC6/GGAACGATGAGCCTCCAAC-3’

PCR_F 5’-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACG CTCTTC-3’

PE_t_N2 5’-ACACTCTTTCCCT ACACGACGCTCTTCCGA TCTN*N-3’

PE_b_A 5’-/5Phos/AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAG-3’

SolSeq6.3.3 (Read1) 5’-TACACCACTCTCAGATGTTATCGAGGTCCGAC -3’

SolSeqV6.3.2r(Read2) 5’-GCTAGGAACGATGAGCCTCCAAC-3’

AmpR6.3IndSeq(IndexRead) 5’-GTTGGAGGCTCATCGTTCCTAGC-3’

Primers used with LC Sciences Probes

eMIP_CA1_F 5’- TGCCTAGGACCGGATCAACT-3’

eMIP_CA1_R 5’- GAGCTTCGGTTCACGCAATG-3’

CP-2-FA 5’-GCACGATCCGACGGTAGTGT-3’

CP-2-RA 5’-CCGTAATCGGGAAGCTGAAG-3’

CA-2-FA.Indx7Sol 5’-CAAGCAGAAGACGGCATACGAGATGATCTGCGGTCTGCCATCCGACGGTAGTGT-3’

CA-2-FA.Indx45Sol 5’-CAAGCAGAAGACGGCATACGAGATCGTAGTCGGTCTGCCATCCGACGGTAGTGT-3’

CA-2-FA.Indx76Sol 5’-CAAGCAGAAGACGGCATACGAGATAATAGGCGGTCTGCCATCCGACGGTAGTGT-3’

CA-2-RA.Sol 5’- AATGATACGGCGACCACCGAGATCTACACGCCTATCGGGAAGCTGAAG-3’

Switch.CA-2-FA.Sol 5’- AATGATACGGCGACCACCGAGATCTACACGCCTATCCGACGGTAGTGT-3’

Switch.CA-2-RA.Ind7Sol 5’- CAAGCAGAAGACGGCATACGAGATGATCTGCGGTCTGCCATCGGGAAGCTGAAG-3’

Switch.CA-2-RA.Ind45Sol 5’- CAAGCAGAAGACGGCATACGAGATCGTAGTCGGTCTGCCATCGGGAAGCTGAAG-3’

Switch.CA-2-RA.Ind76Sol 5’- CAAGCAGAAGACGGCATACGAGATAATAGGCGGTCTGCCATCGGGAAGCTGAAG-3’

CP-2-SeqRead1.x (Read1) 5’-TACACGCCTATCGGGAAGCTGAAG-3’

CP-2-IndSeq.x (IndexRead) 5’-ACACTACCGTCGGATGGCAGACCG-3’

CP-2-SeqRead1.y (Read1) 5’-TACACGCCTATCCGACGGTAGTGT-3’

CP-2-IndSeq.y (IndexRead) 5’-CTTCAGCTTCCCGATGGCAGACCG-3’

* Indicates a phosphorothioate bond


15

Supplementary Table 4. Sequences of multiplexed barcoded primers used in the library-free protocol.

Barcode ID Barcode Primer

Ind1 CGTGAT CAAGCAGAAGACGGCATACGAGATCGTGATGCTAGGAACGATGAGCCTCCAAC

Ind2 ACATCG CAAGCAGAAGACGGCATACGAGATACATCGGCTAGGAACGATGAGCCTCCAAC

Ind3 GCCTAA CAAGCAGAAGACGGCATACGAGATGCCTAAGCTAGGAACGATGAGCCTCCAAC

Ind4 TGGTCA CAAGCAGAAGACGGCATACGAGATTGGTCAGCTAGGAACGATGAGCCTCCAAC

Ind5 CACTGT CAAGCAGAAGACGGCATACGAGATCACTGTGCTAGGAACGATGAGCCTCCAAC

Ind6 ATTGGC CAAGCAGAAGACGGCATACGAGATATTGGCGCTAGGAACGATGAGCCTCCAAC

Ind7 GATCTG CAAGCAGAAGACGGCATACGAGATGATCTGGCTAGGAACGATGAGCCTCCAAC

Ind8 TCAAGT CAAGCAGAAGACGGCATACGAGATTCAAGTGCTAGGAACGATGAGCCTCCAAC

Ind9 CTGATC CAAGCAGAAGACGGCATACGAGATCTGATCGCTAGGAACGATGAGCCTCCAAC

Ind10 AAGCTA CAAGCAGAAGACGGCATACGAGATAAGCTAGCTAGGAACGATGAGCCTCCAAC

Ind11 GTAGCC CAAGCAGAAGACGGCATACGAGATGTAGCCGCTAGGAACGATGAGCCTCCAAC

Ind12 TACAAG CAAGCAGAAGACGGCATACGAGATTACAAGGCTAGGAACGATGAGCCTCCAAC

Ind13 TCATGG CAAGCAGAAGACGGCATACGAGATTCATGGGCTAGGAACGATGAGCCTCCAAC

Ind14 TGTCTT CAAGCAGAAGACGGCATACGAGATTGTCTTGCTAGGAACGATGAGCCTCCAAC

Ind15 AGGAAG CAAGCAGAAGACGGCATACGAGATAGGAAGGCTAGGAACGATGAGCCTCCAAC

Ind16 AACCCC CAAGCAGAAGACGGCATACGAGATAACCCCGCTAGGAACGATGAGCCTCCAAC

Ind17 GATGAG CAAGCAGAAGACGGCATACGAGATGATGAGGCTAGGAACGATGAGCCTCCAAC

Ind18 TGAACT CAAGCAGAAGACGGCATACGAGATTGAACTGCTAGGAACGATGAGCCTCCAAC

Ind19 TGCGTC CAAGCAGAAGACGGCATACGAGATTGCGTCGCTAGGAACGATGAGCCTCCAAC

Ind20 GACAGG CAAGCAGAAGACGGCATACGAGATGACAGGGCTAGGAACGATGAGCCTCCAAC

Ind21 GGGTTG CAAGCAGAAGACGGCATACGAGATGGGTTGGCTAGGAACGATGAGCCTCCAAC

Ind22 TCCGAG CAAGCAGAAGACGGCATACGAGATTCCGAGGCTAGGAACGATGAGCCTCCAAC

Ind23 TTTCGA CAAGCAGAAGACGGCATACGAGATTTTCGAGCTAGGAACGATGAGCCTCCAAC

Ind24 GCGAAT CAAGCAGAAGACGGCATACGAGATGCGAATGCTAGGAACGATGAGCCTCCAAC

Ind25 GCAGTA CAAGCAGAAGACGGCATACGAGATGCAGTAGCTAGGAACGATGAGCCTCCAAC

Ind26 TCACGA CAAGCAGAAGACGGCATACGAGATTCACGAGCTAGGAACGATGAGCCTCCAAC

Ind27 CGCGTA CAAGCAGAAGACGGCATACGAGATCGCGTAGCTAGGAACGATGAGCCTCCAAC

Ind28 GCACCT CAAGCAGAAGACGGCATACGAGATGCACCTGCTAGGAACGATGAGCCTCCAAC

Ind29 GTTCGT CAAGCAGAAGACGGCATACGAGATGTTCGTGCTAGGAACGATGAGCCTCCAAC

Ind30 CACTAA CAAGCAGAAGACGGCATACGAGATCACTAAGCTAGGAACGATGAGCCTCCAAC

Ind31 GTGGTG CAAGCAGAAGACGGCATACGAGATGTGGTGGCTAGGAACGATGAGCCTCCAAC

Ind32 CCTTGC CAAGCAGAAGACGGCATACGAGATCCTTGCGCTAGGAACGATGAGCCTCCAAC

Ind33 GTTGCG CAAGCAGAAGACGGCATACGAGATGTTGCGGCTAGGAACGATGAGCCTCCAAC

Ind34 TCAGTT CAAGCAGAAGACGGCATACGAGATTCAGTTGCTAGGAACGATGAGCCTCCAAC

Ind35 CCCGAT CAAGCAGAAGACGGCATACGAGATCCCGATGCTAGGAACGATGAGCCTCCAAC

Ind36 TGCTTG CAAGCAGAAGACGGCATACGAGATTGCTTGGCTAGGAACGATGAGCCTCCAAC

Ind37 TGTAGC CAAGCAGAAGACGGCATACGAGATTGTAGCGCTAGGAACGATGAGCCTCCAAC

Ind38 GCGTGA CAAGCAGAAGACGGCATACGAGATGCGTGAGCTAGGAACGATGAGCCTCCAAC

Ind39 CTCTGG CAAGCAGAAGACGGCATACGAGATCTCTGGGCTAGGAACGATGAGCCTCCAAC

Ind40 CCGTCA CAAGCAGAAGACGGCATACGAGATCCGTCAGCTAGGAACGATGAGCCTCCAAC

Ind41 GTTCCC CAAGCAGAAGACGGCATACGAGATGTTCCCGCTAGGAACGATGAGCCTCCAAC


16

Ind42 CTTTTC CAAGCAGAAGACGGCATACGAGATCTTTTCGCTAGGAACGATGAGCCTCCAAC

Ind43 GGCACT CAAGCAGAAGACGGCATACGAGATGGCACTGCTAGGAACGATGAGCCTCCAAC

Ind44 GGATGC CAAGCAGAAGACGGCATACGAGATGGATGCGCTAGGAACGATGAGCCTCCAAC

Ind45 CGTAGT CAAGCAGAAGACGGCATACGAGATCGTAGTGCTAGGAACGATGAGCCTCCAAC

Ind46 GAAATG CAAGCAGAAGACGGCATACGAGATGAAATGGCTAGGAACGATGAGCCTCCAAC

Ind47 GGAGAG CAAGCAGAAGACGGCATACGAGATGGAGAGGCTAGGAACGATGAGCCTCCAAC

Ind48 TAACGT CAAGCAGAAGACGGCATACGAGATTAACGTGCTAGGAACGATGAGCCTCCAAC

Ind49 ACACAG CAAGCAGAAGACGGCATACGAGATACACAGGCTAGGAACGATGAGCCTCCAAC

Ind50 AAAGGT CAAGCAGAAGACGGCATACGAGATAAAGGTGCTAGGAACGATGAGCCTCCAAC

Ind51 GCGATA CAAGCAGAAGACGGCATACGAGATGCGATAGCTAGGAACGATGAGCCTCCAAC

Ind52 CGTGTC CAAGCAGAAGACGGCATACGAGATCGTGTCGCTAGGAACGATGAGCCTCCAAC

Ind53 GTAGAA CAAGCAGAAGACGGCATACGAGATGTAGAAGCTAGGAACGATGAGCCTCCAAC

Ind54 GGACGT CAAGCAGAAGACGGCATACGAGATGGACGTGCTAGGAACGATGAGCCTCCAAC

Ind55 AGTCGA CAAGCAGAAGACGGCATACGAGATAGTCGAGCTAGGAACGATGAGCCTCCAAC

Ind56 GTCTGA CAAGCAGAAGACGGCATACGAGATGTCTGAGCTAGGAACGATGAGCCTCCAAC

Ind57 GAAGGA CAAGCAGAAGACGGCATACGAGATGAAGGAGCTAGGAACGATGAGCCTCCAAC

Ind58 ATGCTG CAAGCAGAAGACGGCATACGAGATATGCTGGCTAGGAACGATGAGCCTCCAAC

Ind59 TCTATC CAAGCAGAAGACGGCATACGAGATTCTATCGCTAGGAACGATGAGCCTCCAAC

Ind60 ATCTGT CAAGCAGAAGACGGCATACGAGATATCTGTGCTAGGAACGATGAGCCTCCAAC

Ind61 ATAGAG CAAGCAGAAGACGGCATACGAGATATAGAGGCTAGGAACGATGAGCCTCCAAC

Ind62 GCTAAA CAAGCAGAAGACGGCATACGAGATGCTAAAGCTAGGAACGATGAGCCTCCAAC

Ind63 ACCAGG CAAGCAGAAGACGGCATACGAGATACCAGGGCTAGGAACGATGAGCCTCCAAC

Ind64 CCAACT CAAGCAGAAGACGGCATACGAGATCCAACTGCTAGGAACGATGAGCCTCCAAC

Ind65 AAGGAA CAAGCAGAAGACGGCATACGAGATAAGGAAGCTAGGAACGATGAGCCTCCAAC

Ind66 CCTCCA CAAGCAGAAGACGGCATACGAGATCCTCCAGCTAGGAACGATGAGCCTCCAAC

Ind67 CACGTC CAAGCAGAAGACGGCATACGAGATCACGTCGCTAGGAACGATGAGCCTCCAAC

Ind68 CATAAC CAAGCAGAAGACGGCATACGAGATCATAACGCTAGGAACGATGAGCCTCCAAC

Ind69 CCATAT CAAGCAGAAGACGGCATACGAGATCCATATGCTAGGAACGATGAGCCTCCAAC

Ind70 GAAGTC CAAGCAGAAGACGGCATACGAGATGAAGTCGCTAGGAACGATGAGCCTCCAAC

Ind71 CAAAGA CAAGCAGAAGACGGCATACGAGATCAAAGAGCTAGGAACGATGAGCCTCCAAC

Ind72 TGGCAG CAAGCAGAAGACGGCATACGAGATTGGCAGGCTAGGAACGATGAGCCTCCAAC

Ind73 GAGTCC CAAGCAGAAGACGGCATACGAGATGAGTCCGCTAGGAACGATGAGCCTCCAAC

Ind74 TCGCCA CAAGCAGAAGACGGCATACGAGATTCGCCAGCTAGGAACGATGAGCCTCCAAC

Ind75 AAGTCG CAAGCAGAAGACGGCATACGAGATAAGTCGGCTAGGAACGATGAGCCTCCAAC

Ind76 AATAGG CAAGCAGAAGACGGCATACGAGATAATAGGGCTAGGAACGATGAGCCTCCAAC

Ind77 ACCCGT CAAGCAGAAGACGGCATACGAGATACCCGTGCTAGGAACGATGAGCCTCCAAC

Ind78 AACACG CAAGCAGAAGACGGCATACGAGATAACACGGCTAGGAACGATGAGCCTCCAAC

Ind79 GCTTGG CAAGCAGAAGACGGCATACGAGATGCTTGGGCTAGGAACGATGAGCCTCCAAC

Ind80 TTACCA CAAGCAGAAGACGGCATACGAGATTTACCAGCTAGGAACGATGAGCCTCCAAC

Ind81 CCAGGT CAAGCAGAAGACGGCATACGAGATCCAGGTGCTAGGAACGATGAGCCTCCAAC

Ind82 CGTTTG CAAGCAGAAGACGGCATACGAGATCGTTTGGCTAGGAACGATGAGCCTCCAAC

Ind83 GACCAC CAAGCAGAAGACGGCATACGAGATGACCACGCTAGGAACGATGAGCCTCCAAC

Ind84 ACAAGA CAAGCAGAAGACGGCATACGAGATACAAGAGCTAGGAACGATGAGCCTCCAAC


17

Ind85 ACCGCA CAAGCAGAAGACGGCATACGAGATACCGCAGCTAGGAACGATGAGCCTCCAAC

Ind86 TGGGTA CAAGCAGAAGACGGCATACGAGATTGGGTAGCTAGGAACGATGAGCCTCCAAC

Ind87 ATTCCG CAAGCAGAAGACGGCATACGAGATATTCCGGCTAGGAACGATGAGCCTCCAAC

Ind88 GAATGT CAAGCAGAAGACGGCATACGAGATGAATGTGCTAGGAACGATGAGCCTCCAAC

Ind89 GCTGAT CAAGCAGAAGACGGCATACGAGATGCTGATGCTAGGAACGATGAGCCTCCAAC

Ind90 AGTGCT CAAGCAGAAGACGGCATACGAGATAGTGCTGCTAGGAACGATGAGCCTCCAAC

Ind91 CAGGGA CAAGCAGAAGACGGCATACGAGATCAGGGAGCTAGGAACGATGAGCCTCCAAC

Ind92 CATGCG CAAGCAGAAGACGGCATACGAGATCATGCGGCTAGGAACGATGAGCCTCCAAC

Ind93 TGCCTA CAAGCAGAAGACGGCATACGAGATTGCCTAGCTAGGAACGATGAGCCTCCAAC

Ind94 CTATAC CAAGCAGAAGACGGCATACGAGATCTATACGCTAGGAACGATGAGCCTCCAAC

Ind95 CCGAGT CAAGCAGAAGACGGCATACGAGATCCGAGTGCTAGGAACGATGAGCCTCCAAC

Ind96 ACCTGC CAAGCAGAAGACGGCATACGAGATACCTGCGCTAGGAACGATGAGCCTCCAAC

Ind97 CAGGAC CAAGCAGAAGACGGCATACGAGATCAGGACGCTAGGAACGATGAGCCTCCAAC

Ind98 AAGATG CAAGCAGAAGACGGCATACGAGATAAGATGGCTAGGAACGATGAGCCTCCAAC

Ind99 GGCTTC CAAGCAGAAGACGGCATACGAGATGGCTTCGCTAGGAACGATGAGCCTCCAAC

Ind100 GTGCTT CAAGCAGAAGACGGCATACGAGATGTGCTTGCTAGGAACGATGAGCCTCCAAC

Ind101 TGCGCA CAAGCAGAAGACGGCATACGAGATTGCGCAGCTAGGAACGATGAGCCTCCAAC

Ind102 ACTAGC CAAGCAGAAGACGGCATACGAGATACTAGCGCTAGGAACGATGAGCCTCCAAC

Ind103 TCGAAG CAAGCAGAAGACGGCATACGAGATTCGAAGGCTAGGAACGATGAGCCTCCAAC

Ind104 AGACTA CAAGCAGAAGACGGCATACGAGATAGACTAGCTAGGAACGATGAGCCTCCAAC

Ind105 CGGGTT CAAGCAGAAGACGGCATACGAGATCGGGTTGCTAGGAACGATGAGCCTCCAAC

Ind106 TGACTG CAAGCAGAAGACGGCATACGAGATTGACTGGCTAGGAACGATGAGCCTCCAAC

Ind107 TTGTGT CAAGCAGAAGACGGCATACGAGATTTGTGTGCTAGGAACGATGAGCCTCCAAC

Ind108 TCGCTG CAAGCAGAAGACGGCATACGAGATTCGCTGGCTAGGAACGATGAGCCTCCAAC

Ind109 GATACA CAAGCAGAAGACGGCATACGAGATGATACAGCTAGGAACGATGAGCCTCCAAC

Ind110 TCCTTA CAAGCAGAAGACGGCATACGAGATTCCTTAGCTAGGAACGATGAGCCTCCAAC

Ind111 CGATTT CAAGCAGAAGACGGCATACGAGATCGATTTGCTAGGAACGATGAGCCTCCAAC

Ind112 TTACGG CAAGCAGAAGACGGCATACGAGATTTACGGGCTAGGAACGATGAGCCTCCAAC

Ind113 TGTGAC CAAGCAGAAGACGGCATACGAGATTGTGACGCTAGGAACGATGAGCCTCCAAC

Ind114 TTGGAA CAAGCAGAAGACGGCATACGAGATTTGGAAGCTAGGAACGATGAGCCTCCAAC

Ind115 ACCATA CAAGCAGAAGACGGCATACGAGATACCATAGCTAGGAACGATGAGCCTCCAAC

Ind116 GTCGAG CAAGCAGAAGACGGCATACGAGATGTCGAGGCTAGGAACGATGAGCCTCCAAC

Ind117 ACTGCC CAAGCAGAAGACGGCATACGAGATACTGCCGCTAGGAACGATGAGCCTCCAAC

Ind118 TCGGGT CAAGCAGAAGACGGCATACGAGATTCGGGTGCTAGGAACGATGAGCCTCCAAC

Ind119 GGCATG CAAGCAGAAGACGGCATACGAGATGGCATGGCTAGGAACGATGAGCCTCCAAC

Ind120 GTTTCT CAAGCAGAAGACGGCATACGAGATGTTTCTGCTAGGAACGATGAGCCTCCAAC

Ind121 ACCAAT CAAGCAGAAGACGGCATACGAGATACCAATGCTAGGAACGATGAGCCTCCAAC

Ind122 ATGCAT CAAGCAGAAGACGGCATACGAGATATGCATGCTAGGAACGATGAGCCTCCAAC

Ind123 TACGGC CAAGCAGAAGACGGCATACGAGATTACGGCGCTAGGAACGATGAGCCTCCAAC

Ind124 AGTCCC CAAGCAGAAGACGGCATACGAGATAGTCCCGCTAGGAACGATGAGCCTCCAAC

Ind125 CTGCAG CAAGCAGAAGACGGCATACGAGATCTGCAGGCTAGGAACGATGAGCCTCCAAC

Ind126 CTGTTG CAAGCAGAAGACGGCATACGAGATCTGTTGGCTAGGAACGATGAGCCTCCAAC

Ind127 CGGACA CAAGCAGAAGACGGCATACGAGATCGGACAGCTAGGAACGATGAGCCTCCAAC


18

Ind128 TAAGCG CAAGCAGAAGACGGCATACGAGATTAAGCGGCTAGGAACGATGAGCCTCCAAC

Ind129 GAGAGT CAAGCAGAAGACGGCATACGAGATGAGAGTGCTAGGAACGATGAGCCTCCAAC

Ind130 TACCCG CAAGCAGAAGACGGCATACGAGATTACCCGGCTAGGAACGATGAGCCTCCAAC

Ind131 TTCGTA CAAGCAGAAGACGGCATACGAGATTTCGTAGCTAGGAACGATGAGCCTCCAAC

Ind132 AAAGTG CAAGCAGAAGACGGCATACGAGATAAAGTGGCTAGGAACGATGAGCCTCCAAC

Ind133 TTTGGT CAAGCAGAAGACGGCATACGAGATTTTGGTGCTAGGAACGATGAGCCTCCAAC

Ind134 GTCCCT CAAGCAGAAGACGGCATACGAGATGTCCCTGCTAGGAACGATGAGCCTCCAAC

Ind135 TAGCTT CAAGCAGAAGACGGCATACGAGATTAGCTTGCTAGGAACGATGAGCCTCCAAC

Ind136 GCACTG CAAGCAGAAGACGGCATACGAGATGCACTGGCTAGGAACGATGAGCCTCCAAC

Ind137 ACTATG CAAGCAGAAGACGGCATACGAGATACTATGGCTAGGAACGATGAGCCTCCAAC

Ind138 GAACTT CAAGCAGAAGACGGCATACGAGATGAACTTGCTAGGAACGATGAGCCTCCAAC

Ind139 TTTGAG CAAGCAGAAGACGGCATACGAGATTTTGAGGCTAGGAACGATGAGCCTCCAAC

Ind140 AGGCCT CAAGCAGAAGACGGCATACGAGATAGGCCTGCTAGGAACGATGAGCCTCCAAC

Ind141 ACACGC CAAGCAGAAGACGGCATACGAGATACACGCGCTAGGAACGATGAGCCTCCAAC

Ind142 TTGTCG CAAGCAGAAGACGGCATACGAGATTTGTCGGCTAGGAACGATGAGCCTCCAAC

Ind143 TCTCAC CAAGCAGAAGACGGCATACGAGATTCTCACGCTAGGAACGATGAGCCTCCAAC

Ind144 TAGACC CAAGCAGAAGACGGCATACGAGATTAGACCGCTAGGAACGATGAGCCTCCAAC

Ind145 CCTAAG CAAGCAGAAGACGGCATACGAGATCCTAAGGCTAGGAACGATGAGCCTCCAAC

Ind146 GTATCG CAAGCAGAAGACGGCATACGAGATGTATCGGCTAGGAACGATGAGCCTCCAAC

Ind147 TCCAGA CAAGCAGAAGACGGCATACGAGATTCCAGAGCTAGGAACGATGAGCCTCCAAC

Ind148 AGGTGA CAAGCAGAAGACGGCATACGAGATAGGTGAGCTAGGAACGATGAGCCTCCAAC

Ind149 CCCATC CAAGCAGAAGACGGCATACGAGATCCCATCGCTAGGAACGATGAGCCTCCAAC

Ind150 TTGCAC CAAGCAGAAGACGGCATACGAGATTTGCACGCTAGGAACGATGAGCCTCCAAC

Ind151 AACTCA CAAGCAGAAGACGGCATACGAGATAACTCAGCTAGGAACGATGAGCCTCCAAC

Ind152 CGTATA CAAGCAGAAGACGGCATACGAGATCGTATAGCTAGGAACGATGAGCCTCCAAC

Ind153 AGCGAA CAAGCAGAAGACGGCATACGAGATAGCGAAGCTAGGAACGATGAGCCTCCAAC

Ind154 ACGGCT CAAGCAGAAGACGGCATACGAGATACGGCTGCTAGGAACGATGAGCCTCCAAC

Ind155 AGTGAG CAAGCAGAAGACGGCATACGAGATAGTGAGGCTAGGAACGATGAGCCTCCAAC

Ind156 TTTCTC CAAGCAGAAGACGGCATACGAGATTTTCTCGCTAGGAACGATGAGCCTCCAAC

Ind157 GCTCTA CAAGCAGAAGACGGCATACGAGATGCTCTAGCTAGGAACGATGAGCCTCCAAC

Ind158 ACTTGA CAAGCAGAAGACGGCATACGAGATACTTGAGCTAGGAACGATGAGCCTCCAAC

Ind159 CGGTTC CAAGCAGAAGACGGCATACGAGATCGGTTCGCTAGGAACGATGAGCCTCCAAC

Ind160 CATCAT CAAGCAGAAGACGGCATACGAGATCATCATGCTAGGAACGATGAGCCTCCAAC

Ind161 CAAACG CAAGCAGAAGACGGCATACGAGATCAAACGGCTAGGAACGATGAGCCTCCAAC

Ind162 CTATGT CAAGCAGAAGACGGCATACGAGATCTATGTGCTAGGAACGATGAGCCTCCAAC

Ind163 AGCGTT CAAGCAGAAGACGGCATACGAGATAGCGTTGCTAGGAACGATGAGCCTCCAAC

Ind164 AAGACT CAAGCAGAAGACGGCATACGAGATAAGACTGCTAGGAACGATGAGCCTCCAAC

Ind165 CGATAA CAAGCAGAAGACGGCATACGAGATCGATAAGCTAGGAACGATGAGCCTCCAAC

Ind166 CGGCTA CAAGCAGAAGACGGCATACGAGATCGGCTAGCTAGGAACGATGAGCCTCCAAC

Ind167 TATACG CAAGCAGAAGACGGCATACGAGATTATACGGCTAGGAACGATGAGCCTCCAAC

Ind168 GAAACC CAAGCAGAAGACGGCATACGAGATGAAACCGCTAGGAACGATGAGCCTCCAAC

Ind169 GAACCG CAAGCAGAAGACGGCATACGAGATGAACCGGCTAGGAACGATGAGCCTCCAAC

Ind170 TTAGGC CAAGCAGAAGACGGCATACGAGATTTAGGCGCTAGGAACGATGAGCCTCCAAC


19

Ind171 GCCTTT CAAGCAGAAGACGGCATACGAGATGCCTTTGCTAGGAACGATGAGCCTCCAAC

Ind172 GTTGGA CAAGCAGAAGACGGCATACGAGATGTTGGAGCTAGGAACGATGAGCCTCCAAC

Ind173 GTACGC CAAGCAGAAGACGGCATACGAGATGTACGCGCTAGGAACGATGAGCCTCCAAC

Ind174 TGGAAC CAAGCAGAAGACGGCATACGAGATTGGAACGCTAGGAACGATGAGCCTCCAAC

Ind175 AACAGA CAAGCAGAAGACGGCATACGAGATAACAGAGCTAGGAACGATGAGCCTCCAAC

Ind176 AAGCAC CAAGCAGAAGACGGCATACGAGATAAGCACGCTAGGAACGATGAGCCTCCAAC

Ind177 ATGGTA CAAGCAGAAGACGGCATACGAGATATGGTAGCTAGGAACGATGAGCCTCCAAC

Ind178 TCGTTC CAAGCAGAAGACGGCATACGAGATTCGTTCGCTAGGAACGATGAGCCTCCAAC

Ind179 CTTCTA CAAGCAGAAGACGGCATACGAGATCTTCTAGCTAGGAACGATGAGCCTCCAAC

Ind180 TGGGAT CAAGCAGAAGACGGCATACGAGATTGGGATGCTAGGAACGATGAGCCTCCAAC

Ind181 ATCGCC CAAGCAGAAGACGGCATACGAGATATCGCCGCTAGGAACGATGAGCCTCCAAC

Ind182 AGTTGG CAAGCAGAAGACGGCATACGAGATAGTTGGGCTAGGAACGATGAGCCTCCAAC

Ind183 AGCTCT CAAGCAGAAGACGGCATACGAGATAGCTCTGCTAGGAACGATGAGCCTCCAAC

Ind184 GACGGT CAAGCAGAAGACGGCATACGAGATGACGGTGCTAGGAACGATGAGCCTCCAAC

Ind185 CTCAGC CAAGCAGAAGACGGCATACGAGATCTCAGCGCTAGGAACGATGAGCCTCCAAC

Ind186 CTTAGG CAAGCAGAAGACGGCATACGAGATCTTAGGGCTAGGAACGATGAGCCTCCAAC

Ind187 CGACAG CAAGCAGAAGACGGCATACGAGATCGACAGGCTAGGAACGATGAGCCTCCAAC

Ind188 ACATGT CAAGCAGAAGACGGCATACGAGATACATGTGCTAGGAACGATGAGCCTCCAAC

Ind189 ATACGA CAAGCAGAAGACGGCATACGAGATATACGAGCTAGGAACGATGAGCCTCCAAC

Ind190 GAGCAT CAAGCAGAAGACGGCATACGAGATGAGCATGCTAGGAACGATGAGCCTCCAAC

Ind191 ATCCTA CAAGCAGAAGACGGCATACGAGATATCCTAGCTAGGAACGATGAGCCTCCAAC

Ind192 ACGTAA CAAGCAGAAGACGGCATACGAGATACGTAAGCTAGGAACGATGAGCCTCCAAC

Ind193 GGAAAC CAAGCAGAAGACGGCATACGAGATGGAAACGCTAGGAACGATGAGCCTCCAAC

Ind194 AGCAAC CAAGCAGAAGACGGCATACGAGATAGCAACGCTAGGAACGATGAGCCTCCAAC

Ind195 CTAGTG CAAGCAGAAGACGGCATACGAGATCTAGTGGCTAGGAACGATGAGCCTCCAAC

Ind196 CCGCTT CAAGCAGAAGACGGCATACGAGATCCGCTTGCTAGGAACGATGAGCCTCCAAC

Ind197 CCAGCA CAAGCAGAAGACGGCATACGAGATCCAGCAGCTAGGAACGATGAGCCTCCAAC

Ind198 ATACTC CAAGCAGAAGACGGCATACGAGATATACTCGCTAGGAACGATGAGCCTCCAAC

Ind199 GGTGAA CAAGCAGAAGACGGCATACGAGATGGTGAAGCTAGGAACGATGAGCCTCCAAC

Ind200 CTCTAT CAAGCAGAAGACGGCATACGAGATCTCTATGCTAGGAACGATGAGCCTCCAAC

Ind201 GACATA CAAGCAGAAGACGGCATACGAGATGACATAGCTAGGAACGATGAGCCTCCAAC

Ind202 GGATCT CAAGCAGAAGACGGCATACGAGATGGATCTGCTAGGAACGATGAGCCTCCAAC

Ind203 AACGAG CAAGCAGAAGACGGCATACGAGATAACGAGGCTAGGAACGATGAGCCTCCAAC

Ind204 CACCTG CAAGCAGAAGACGGCATACGAGATCACCTGGCTAGGAACGATGAGCCTCCAAC

Ind205 CATGAA CAAGCAGAAGACGGCATACGAGATCATGAAGCTAGGAACGATGAGCCTCCAAC

Ind206 CGTGGA CAAGCAGAAGACGGCATACGAGATCGTGGAGCTAGGAACGATGAGCCTCCAAC

Ind207 AGAAGG CAAGCAGAAGACGGCATACGAGATAGAAGGGCTAGGAACGATGAGCCTCCAAC

Ind208 ATCCAG CAAGCAGAAGACGGCATACGAGATATCCAGGCTAGGAACGATGAGCCTCCAAC

Ind209 TTCCTG CAAGCAGAAGACGGCATACGAGATTTCCTGGCTAGGAACGATGAGCCTCCAAC

Ind210 CAACAA CAAGCAGAAGACGGCATACGAGATCAACAAGCTAGGAACGATGAGCCTCCAAC

Ind211 CCTGTT CAAGCAGAAGACGGCATACGAGATCCTGTTGCTAGGAACGATGAGCCTCCAAC

Ind212 CTCGTT CAAGCAGAAGACGGCATACGAGATCTCGTTGCTAGGAACGATGAGCCTCCAAC

Ind213 CTGAGA CAAGCAGAAGACGGCATACGAGATCTGAGAGCTAGGAACGATGAGCCTCCAAC


20

Ind214 CGCTCA CAAGCAGAAGACGGCATACGAGATCGCTCAGCTAGGAACGATGAGCCTCCAAC

Ind215 TACCAA CAAGCAGAAGACGGCATACGAGATTACCAAGCTAGGAACGATGAGCCTCCAAC

Ind216 CGCAAG CAAGCAGAAGACGGCATACGAGATCGCAAGGCTAGGAACGATGAGCCTCCAAC

Ind217 TACTGA CAAGCAGAAGACGGCATACGAGATTACTGAGCTAGGAACGATGAGCCTCCAAC

Ind218 TCCACT CAAGCAGAAGACGGCATACGAGATTCCACTGCTAGGAACGATGAGCCTCCAAC

Ind219 ATCGTG CAAGCAGAAGACGGCATACGAGATATCGTGGCTAGGAACGATGAGCCTCCAAC

Ind220 TAGGCA CAAGCAGAAGACGGCATACGAGATTAGGCAGCTAGGAACGATGAGCCTCCAAC

Ind221 CAAGAG CAAGCAGAAGACGGCATACGAGATCAAGAGGCTAGGAACGATGAGCCTCCAAC

Ind222 ACGACA CAAGCAGAAGACGGCATACGAGATACGACAGCTAGGAACGATGAGCCTCCAAC

Ind223 GGTTCC CAAGCAGAAGACGGCATACGAGATGGTTCCGCTAGGAACGATGAGCCTCCAAC

Ind224 TCTTAG CAAGCAGAAGACGGCATACGAGATTCTTAGGCTAGGAACGATGAGCCTCCAAC

Ind225 AGAGGA CAAGCAGAAGACGGCATACGAGATAGAGGAGCTAGGAACGATGAGCCTCCAAC

Ind226 AAAGCA CAAGCAGAAGACGGCATACGAGATAAAGCAGCTAGGAACGATGAGCCTCCAAC

Ind227 AGTCAT CAAGCAGAAGACGGCATACGAGATAGTCATGCTAGGAACGATGAGCCTCCAAC

Ind228 TGCAGT CAAGCAGAAGACGGCATACGAGATTGCAGTGCTAGGAACGATGAGCCTCCAAC

Ind229 TGTTCT CAAGCAGAAGACGGCATACGAGATTGTTCTGCTAGGAACGATGAGCCTCCAAC

Ind230 TATCGC CAAGCAGAAGACGGCATACGAGATTATCGCGCTAGGAACGATGAGCCTCCAAC

Ind231 GCATCA CAAGCAGAAGACGGCATACGAGATGCATCAGCTAGGAACGATGAGCCTCCAAC

Ind232 CTCCGT CAAGCAGAAGACGGCATACGAGATCTCCGTGCTAGGAACGATGAGCCTCCAAC

Ind233 TAAGAC CAAGCAGAAGACGGCATACGAGATTAAGACGCTAGGAACGATGAGCCTCCAAC

Ind234 GAGGCT CAAGCAGAAGACGGCATACGAGATGAGGCTGCTAGGAACGATGAGCCTCCAAC

Ind235 TGATTC CAAGCAGAAGACGGCATACGAGATTGATTCGCTAGGAACGATGAGCCTCCAAC

Ind236 GTCCAA CAAGCAGAAGACGGCATACGAGATGTCCAAGCTAGGAACGATGAGCCTCCAAC

Ind237 ACTCTC CAAGCAGAAGACGGCATACGAGATACTCTCGCTAGGAACGATGAGCCTCCAAC

Ind238 TCAACG CAAGCAGAAGACGGCATACGAGATTCAACGGCTAGGAACGATGAGCCTCCAAC

Ind239 GGGTAC CAAGCAGAAGACGGCATACGAGATGGGTACGCTAGGAACGATGAGCCTCCAAC

Ind240 ACGATT CAAGCAGAAGACGGCATACGAGATACGATTGCTAGGAACGATGAGCCTCCAAC

Ind241 AACTTG CAAGCAGAAGACGGCATACGAGATAACTTGGCTAGGAACGATGAGCCTCCAAC

Ind242 AACGTA CAAGCAGAAGACGGCATACGAGATAACGTAGCTAGGAACGATGAGCCTCCAAC

Ind243 ACCCAC CAAGCAGAAGACGGCATACGAGATACCCACGCTAGGAACGATGAGCCTCCAAC

Ind244 CGGTCT CAAGCAGAAGACGGCATACGAGATCGGTCTGCTAGGAACGATGAGCCTCCAAC

Ind245 TTCTAG CAAGCAGAAGACGGCATACGAGATTTCTAGGCTAGGAACGATGAGCCTCCAAC

Ind246 GGTGTG CAAGCAGAAGACGGCATACGAGATGGTGTGGCTAGGAACGATGAGCCTCCAAC

Ind247 ACCACC CAAGCAGAAGACGGCATACGAGATACCACCGCTAGGAACGATGAGCCTCCAAC

Ind248 GACTGC CAAGCAGAAGACGGCATACGAGATGACTGCGCTAGGAACGATGAGCCTCCAAC

Ind249 CTACTT CAAGCAGAAGACGGCATACGAGATCTACTTGCTAGGAACGATGAGCCTCCAAC

Ind250 TAGCGG CAAGCAGAAGACGGCATACGAGATTAGCGGGCTAGGAACGATGAGCCTCCAAC

Ind251 TGTGCG CAAGCAGAAGACGGCATACGAGATTGTGCGGCTAGGAACGATGAGCCTCCAAC

Ind252 CTGGCT CAAGCAGAAGACGGCATACGAGATCTGGCTGCTAGGAACGATGAGCCTCCAAC

Ind253 CGAGAC CAAGCAGAAGACGGCATACGAGATCGAGACGCTAGGAACGATGAGCCTCCAAC

Ind254 GGGATT CAAGCAGAAGACGGCATACGAGATGGGATTGCTAGGAACGATGAGCCTCCAAC

Ind255 TACTCC CAAGCAGAAGACGGCATACGAGATTACTCCGCTAGGAACGATGAGCCTCCAAC

Ind256 GTGGCA CAAGCAGAAGACGGCATACGAGATGTGGCAGCTAGGAACGATGAGCCTCCAAC


21

Ind257 GTCGTC CAAGCAGAAGACGGCATACGAGATGTCGTCGCTAGGAACGATGAGCCTCCAAC

Ind258 GCATTC CAAGCAGAAGACGGCATACGAGATGCATTCGCTAGGAACGATGAGCCTCCAAC

Ind259 GAGCGA CAAGCAGAAGACGGCATACGAGATGAGCGAGCTAGGAACGATGAGCCTCCAAC

Ind260 AGACAC CAAGCAGAAGACGGCATACGAGATAGACACGCTAGGAACGATGAGCCTCCAAC

Ind261 TCTGGG CAAGCAGAAGACGGCATACGAGATTCTGGGGCTAGGAACGATGAGCCTCCAAC

Ind262 GCCAGT CAAGCAGAAGACGGCATACGAGATGCCAGTGCTAGGAACGATGAGCCTCCAAC

Ind263 CCAGTC CAAGCAGAAGACGGCATACGAGATCCAGTCGCTAGGAACGATGAGCCTCCAAC

Ind264 TTGGCC CAAGCAGAAGACGGCATACGAGATTTGGCCGCTAGGAACGATGAGCCTCCAAC

Ind265 GTAACA CAAGCAGAAGACGGCATACGAGATGTAACAGCTAGGAACGATGAGCCTCCAAC

Ind266 ATTACC CAAGCAGAAGACGGCATACGAGATATTACCGCTAGGAACGATGAGCCTCCAAC

Ind267 TCTCCT CAAGCAGAAGACGGCATACGAGATTCTCCTGCTAGGAACGATGAGCCTCCAAC

Ind268 AGATCA CAAGCAGAAGACGGCATACGAGATAGATCAGCTAGGAACGATGAGCCTCCAAC

Ind269 TCCTAT CAAGCAGAAGACGGCATACGAGATTCCTATGCTAGGAACGATGAGCCTCCAAC

Ind270 ACTCGG CAAGCAGAAGACGGCATACGAGATACTCGGGCTAGGAACGATGAGCCTCCAAC

Ind271 TGCCAT CAAGCAGAAGACGGCATACGAGATTGCCATGCTAGGAACGATGAGCCTCCAAC

Ind272 CATCTC CAAGCAGAAGACGGCATACGAGATCATCTCGCTAGGAACGATGAGCCTCCAAC

Ind273 CTTTGA CAAGCAGAAGACGGCATACGAGATCTTTGAGCTAGGAACGATGAGCCTCCAAC

Ind274 TCGCAT CAAGCAGAAGACGGCATACGAGATTCGCATGCTAGGAACGATGAGCCTCCAAC

Ind275 CACACC CAAGCAGAAGACGGCATACGAGATCACACCGCTAGGAACGATGAGCCTCCAAC

Ind276 ACAGAA CAAGCAGAAGACGGCATACGAGATACAGAAGCTAGGAACGATGAGCCTCCAAC

Ind277 ATGTCA CAAGCAGAAGACGGCATACGAGATATGTCAGCTAGGAACGATGAGCCTCCAAC

Ind278 CTGTCC CAAGCAGAAGACGGCATACGAGATCTGTCCGCTAGGAACGATGAGCCTCCAAC

Ind279 CGATCC CAAGCAGAAGACGGCATACGAGATCGATCCGCTAGGAACGATGAGCCTCCAAC

Ind280 TGGAGG CAAGCAGAAGACGGCATACGAGATTGGAGGGCTAGGAACGATGAGCCTCCAAC

Ind281 CGCCAA CAAGCAGAAGACGGCATACGAGATCGCCAAGCTAGGAACGATGAGCCTCCAAC

Ind282 GAATAG CAAGCAGAAGACGGCATACGAGATGAATAGGCTAGGAACGATGAGCCTCCAAC

Ind283 CAACGG CAAGCAGAAGACGGCATACGAGATCAACGGGCTAGGAACGATGAGCCTCCAAC

Ind284 GCGGAA CAAGCAGAAGACGGCATACGAGATGCGGAAGCTAGGAACGATGAGCCTCCAAC

Ind285 TCTGCA CAAGCAGAAGACGGCATACGAGATTCTGCAGCTAGGAACGATGAGCCTCCAAC

Ind286 GATGGC CAAGCAGAAGACGGCATACGAGATGATGGCGCTAGGAACGATGAGCCTCCAAC

Ind287 CCGATG CAAGCAGAAGACGGCATACGAGATCCGATGGCTAGGAACGATGAGCCTCCAAC

Ind288 GATTTC CAAGCAGAAGACGGCATACGAGATGATTTCGCTAGGAACGATGAGCCTCCAAC

Ind289 CCAAAC CAAGCAGAAGACGGCATACGAGATCCAAACGCTAGGAACGATGAGCCTCCAAC

Ind290 AGGATC CAAGCAGAAGACGGCATACGAGATAGGATCGCTAGGAACGATGAGCCTCCAAC

Ind291 CATTCA CAAGCAGAAGACGGCATACGAGATCATTCAGCTAGGAACGATGAGCCTCCAAC

Ind292 AGATTG CAAGCAGAAGACGGCATACGAGATAGATTGGCTAGGAACGATGAGCCTCCAAC

Ind293 CGAAGC CAAGCAGAAGACGGCATACGAGATCGAAGCGCTAGGAACGATGAGCCTCCAAC

Ind294 GGAACG CAAGCAGAAGACGGCATACGAGATGGAACGGCTAGGAACGATGAGCCTCCAAC

Ind295 CGACCA CAAGCAGAAGACGGCATACGAGATCGACCAGCTAGGAACGATGAGCCTCCAAC

Ind296 AGCTTA CAAGCAGAAGACGGCATACGAGATAGCTTAGCTAGGAACGATGAGCCTCCAAC

Ind297 TTCACG CAAGCAGAAGACGGCATACGAGATTTCACGGCTAGGAACGATGAGCCTCCAAC

Ind298 CATTAG CAAGCAGAAGACGGCATACGAGATCATTAGGCTAGGAACGATGAGCCTCCAAC

Ind299 TAGGAG CAAGCAGAAGACGGCATACGAGATTAGGAGGCTAGGAACGATGAGCCTCCAAC


22

Ind300 CTACCG CAAGCAGAAGACGGCATACGAGATCTACCGGCTAGGAACGATGAGCCTCCAAC

Ind301 ATATCC CAAGCAGAAGACGGCATACGAGATATATCCGCTAGGAACGATGAGCCTCCAAC

Ind302 AATGGA CAAGCAGAAGACGGCATACGAGATAATGGAGCTAGGAACGATGAGCCTCCAAC

Ind303 TTCGAC CAAGCAGAAGACGGCATACGAGATTTCGACGCTAGGAACGATGAGCCTCCAAC

Ind304 AACCGG CAAGCAGAAGACGGCATACGAGATAACCGGGCTAGGAACGATGAGCCTCCAAC

Ind305 AAGGCC CAAGCAGAAGACGGCATACGAGATAAGGCCGCTAGGAACGATGAGCCTCCAAC

Ind306 CGTCCT CAAGCAGAAGACGGCATACGAGATCGTCCTGCTAGGAACGATGAGCCTCCAAC

Ind307 AAGAGC CAAGCAGAAGACGGCATACGAGATAAGAGCGCTAGGAACGATGAGCCTCCAAC

Ind308 GTGAAA CAAGCAGAAGACGGCATACGAGATGTGAAAGCTAGGAACGATGAGCCTCCAAC

Ind309 ACGAAC CAAGCAGAAGACGGCATACGAGATACGAACGCTAGGAACGATGAGCCTCCAAC

Ind310 CCGAAA CAAGCAGAAGACGGCATACGAGATCCGAAAGCTAGGAACGATGAGCCTCCAAC

Ind311 CAAGGC CAAGCAGAAGACGGCATACGAGATCAAGGCGCTAGGAACGATGAGCCTCCAAC

Ind312 ATGTGC CAAGCAGAAGACGGCATACGAGATATGTGCGCTAGGAACGATGAGCCTCCAAC

Ind313 CCCTTG CAAGCAGAAGACGGCATACGAGATCCCTTGGCTAGGAACGATGAGCCTCCAAC

Ind314 GAGAAG CAAGCAGAAGACGGCATACGAGATGAGAAGGCTAGGAACGATGAGCCTCCAAC

Ind315 GTAGTT CAAGCAGAAGACGGCATACGAGATGTAGTTGCTAGGAACGATGAGCCTCCAAC

Ind316 TGGTGC CAAGCAGAAGACGGCATACGAGATTGGTGCGCTAGGAACGATGAGCCTCCAAC

Ind317 GACTAT CAAGCAGAAGACGGCATACGAGATGACTATGCTAGGAACGATGAGCCTCCAAC

Ind318 CTCGCA CAAGCAGAAGACGGCATACGAGATCTCGCAGCTAGGAACGATGAGCCTCCAAC

Ind319 TCTTGT CAAGCAGAAGACGGCATACGAGATTCTTGTGCTAGGAACGATGAGCCTCCAAC

Ind320 GCACAA CAAGCAGAAGACGGCATACGAGATGCACAAGCTAGGAACGATGAGCCTCCAAC

Ind321 CCTTTA CAAGCAGAAGACGGCATACGAGATCCTTTAGCTAGGAACGATGAGCCTCCAAC

Ind322 ACGTTG CAAGCAGAAGACGGCATACGAGATACGTTGGCTAGGAACGATGAGCCTCCAAC

Ind323 AAGCGT CAAGCAGAAGACGGCATACGAGATAAGCGTGCTAGGAACGATGAGCCTCCAAC

Ind324 AGGCAA CAAGCAGAAGACGGCATACGAGATAGGCAAGCTAGGAACGATGAGCCTCCAAC

Ind325 TGAGCC CAAGCAGAAGACGGCATACGAGATTGAGCCGCTAGGAACGATGAGCCTCCAAC

Ind326 TGAGAA CAAGCAGAAGACGGCATACGAGATTGAGAAGCTAGGAACGATGAGCCTCCAAC

Ind327 GTACAG CAAGCAGAAGACGGCATACGAGATGTACAGGCTAGGAACGATGAGCCTCCAAC

Ind328 ACGGAG CAAGCAGAAGACGGCATACGAGATACGGAGGCTAGGAACGATGAGCCTCCAAC

Ind329 ACATAC CAAGCAGAAGACGGCATACGAGATACATACGCTAGGAACGATGAGCCTCCAAC

Ind330 CAGCCA CAAGCAGAAGACGGCATACGAGATCAGCCAGCTAGGAACGATGAGCCTCCAAC

Ind331 TGTCAA CAAGCAGAAGACGGCATACGAGATTGTCAAGCTAGGAACGATGAGCCTCCAAC

Ind332 TATTGG CAAGCAGAAGACGGCATACGAGATTATTGGGCTAGGAACGATGAGCCTCCAAC

Ind333 AGACCG CAAGCAGAAGACGGCATACGAGATAGACCGGCTAGGAACGATGAGCCTCCAAC

Ind334 GCCAAC CAAGCAGAAGACGGCATACGAGATGCCAACGCTAGGAACGATGAGCCTCCAAC

Ind335 ATGTAG CAAGCAGAAGACGGCATACGAGATATGTAGGCTAGGAACGATGAGCCTCCAAC

Ind336 CGCTAC CAAGCAGAAGACGGCATACGAGATCGCTACGCTAGGAACGATGAGCCTCCAAC

Ind337 CACTCG CAAGCAGAAGACGGCATACGAGATCACTCGGCTAGGAACGATGAGCCTCCAAC

Ind338 GCTATT CAAGCAGAAGACGGCATACGAGATGCTATTGCTAGGAACGATGAGCCTCCAAC

Ind339 CTCCAC CAAGCAGAAGACGGCATACGAGATCTCCACGCTAGGAACGATGAGCCTCCAAC

Ind340 GTTAGC CAAGCAGAAGACGGCATACGAGATGTTAGCGCTAGGAACGATGAGCCTCCAAC

Ind341 AAACCT CAAGCAGAAGACGGCATACGAGATAAACCTGCTAGGAACGATGAGCCTCCAAC

Ind342 CTAGAT CAAGCAGAAGACGGCATACGAGATCTAGATGCTAGGAACGATGAGCCTCCAAC


23

Ind343 ACCGTC CAAGCAGAAGACGGCATACGAGATACCGTCGCTAGGAACGATGAGCCTCCAAC

Ind344 TCACCC CAAGCAGAAGACGGCATACGAGATTCACCCGCTAGGAACGATGAGCCTCCAAC

Ind345 GAAGAT CAAGCAGAAGACGGCATACGAGATGAAGATGCTAGGAACGATGAGCCTCCAAC

Ind346 TGGCTC CAAGCAGAAGACGGCATACGAGATTGGCTCGCTAGGAACGATGAGCCTCCAAC

Ind347 GGTTAT CAAGCAGAAGACGGCATACGAGATGGTTATGCTAGGAACGATGAGCCTCCAAC

Ind348 ACACTT CAAGCAGAAGACGGCATACGAGATACACTTGCTAGGAACGATGAGCCTCCAAC

Ind349 GGAAGA CAAGCAGAAGACGGCATACGAGATGGAAGAGCTAGGAACGATGAGCCTCCAAC

Ind350 TACGTG CAAGCAGAAGACGGCATACGAGATTACGTGGCTAGGAACGATGAGCCTCCAAC

Ind351 CTTGAC CAAGCAGAAGACGGCATACGAGATCTTGACGCTAGGAACGATGAGCCTCCAAC

Ind352 ATGCCC CAAGCAGAAGACGGCATACGAGATATGCCCGCTAGGAACGATGAGCCTCCAAC

Ind353 ATTCAC CAAGCAGAAGACGGCATACGAGATATTCACGCTAGGAACGATGAGCCTCCAAC

Ind354 GCTTAC CAAGCAGAAGACGGCATACGAGATGCTTACGCTAGGAACGATGAGCCTCCAAC

Ind355 TTCTGC CAAGCAGAAGACGGCATACGAGATTTCTGCGCTAGGAACGATGAGCCTCCAAC

Ind356 TGTATG CAAGCAGAAGACGGCATACGAGATTGTATGGCTAGGAACGATGAGCCTCCAAC

Ind357 TGACGC CAAGCAGAAGACGGCATACGAGATTGACGCGCTAGGAACGATGAGCCTCCAAC

Ind358 GTGTTA CAAGCAGAAGACGGCATACGAGATGTGTTAGCTAGGAACGATGAGCCTCCAAC

Ind359 CATCGA CAAGCAGAAGACGGCATACGAGATCATCGAGCTAGGAACGATGAGCCTCCAAC

Ind360 TGAGGT CAAGCAGAAGACGGCATACGAGATTGAGGTGCTAGGAACGATGAGCCTCCAAC

Ind361 GGTCCA CAAGCAGAAGACGGCATACGAGATGGTCCAGCTAGGAACGATGAGCCTCCAAC

Ind362 TATGCT CAAGCAGAAGACGGCATACGAGATTATGCTGCTAGGAACGATGAGCCTCCAAC

Ind363 TTCATC CAAGCAGAAGACGGCATACGAGATTTCATCGCTAGGAACGATGAGCCTCCAAC

Ind364 CCTCTG CAAGCAGAAGACGGCATACGAGATCCTCTGGCTAGGAACGATGAGCCTCCAAC

Ind365 CCAAGG CAAGCAGAAGACGGCATACGAGATCCAAGGGCTAGGAACGATGAGCCTCCAAC

Ind366 TAATGC CAAGCAGAAGACGGCATACGAGATTAATGCGCTAGGAACGATGAGCCTCCAAC

Ind367 AGGGCA CAAGCAGAAGACGGCATACGAGATAGGGCAGCTAGGAACGATGAGCCTCCAAC

Ind368 TAGAGA CAAGCAGAAGACGGCATACGAGATTAGAGAGCTAGGAACGATGAGCCTCCAAC

Ind369 AGCACA CAAGCAGAAGACGGCATACGAGATAGCACAGCTAGGAACGATGAGCCTCCAAC

Ind370 AGAGTC CAAGCAGAAGACGGCATACGAGATAGAGTCGCTAGGAACGATGAGCCTCCAAC

Ind371 ACTCAA CAAGCAGAAGACGGCATACGAGATACTCAAGCTAGGAACGATGAGCCTCCAAC

Ind372 GTGACG CAAGCAGAAGACGGCATACGAGATGTGACGGCTAGGAACGATGAGCCTCCAAC

Ind373 GGTAGG CAAGCAGAAGACGGCATACGAGATGGTAGGGCTAGGAACGATGAGCCTCCAAC

Ind374 CTGAAT CAAGCAGAAGACGGCATACGAGATCTGAATGCTAGGAACGATGAGCCTCCAAC

Ind375 GTCTAC CAAGCAGAAGACGGCATACGAGATGTCTACGCTAGGAACGATGAGCCTCCAAC

Ind376 TCGGAC CAAGCAGAAGACGGCATACGAGATTCGGACGCTAGGAACGATGAGCCTCCAAC

Ind377 AACTAC CAAGCAGAAGACGGCATACGAGATAACTACGCTAGGAACGATGAGCCTCCAAC

Ind378 AAGGTT CAAGCAGAAGACGGCATACGAGATAAGGTTGCTAGGAACGATGAGCCTCCAAC

Ind379 AGGGAC CAAGCAGAAGACGGCATACGAGATAGGGACGCTAGGAACGATGAGCCTCCAAC

Ind380 ACGCGA CAAGCAGAAGACGGCATACGAGATACGCGAGCTAGGAACGATGAGCCTCCAAC

Ind381 TTGCTA CAAGCAGAAGACGGCATACGAGATTTGCTAGCTAGGAACGATGAGCCTCCAAC

Ind382 ATAACG CAAGCAGAAGACGGCATACGAGATATAACGGCTAGGAACGATGAGCCTCCAAC

Ind383 CCGGTA CAAGCAGAAGACGGCATACGAGATCCGGTAGCTAGGAACGATGAGCCTCCAAC

Ind384 AGCTAG CAAGCAGAAGACGGCATACGAGATAGCTAGGCTAGGAACGATGAGCCTCCAAC


Library-free methylation sequencing with bisulfite padlock ... · Nature Methods Library-free...

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Transcript of Library-free methylation sequencing with bisulfite padlock ... · Nature Methods Library-free...