A convenient system for highly specific and sensitive detection of miRNA expression

Xiangqi Li, Minjie Ni, Chaobao Zhang, Wubin Ma, Yonglian Zhang

SUPPLEMENTAL DATA

A

B

C

miR

-138

miR

-99b＊

miR

-125

a-3p

miR

-291

miR

-292

-5p

miR

-295

miR

-326

miR

-308

5

NO miR

-122

Mar

ker

miR

-138

miR

-99b＊

miR

-125

a-3p

miR

-291

miR

-292

-5p

miR

-295

miR

-326

miR

-308

5

NO miR

-122

Mar

ker

miR

-138

miR

-99b＊

miR

-125

a-3p

miR

-291

miR

-292

-5p

miR

-295

miR

-326

miR

-308

5

NO miR

-122

Mar

ker

Fig. S1 Detection of miRNAs under different conditions by stem-loop PCR.

50bp100bp

50bp100bp

50bp100bp

Fig. S2 The sensitivity of the detection of miR-7578 and distinguishing miR-7578 from its precursor by northern blotting.

Rno-miR-7578

Pre-miR-7578

10 fmol

1 fmol

1 amol

1 zmol

JEG

-3 C

ell

MG

-63

Cell

U-2 O

S cel

l56

37 C

ell

T24 C

ell

Rno-miR-29a by aLHCD

Rno-miR-29a by northern blotting A

B

Fig. S3 Detection of miR-29a in different cell lines by different methods.

Fig. S4 Quantitative detection of miRNA levels using U6 as an internal control by LH-PCR.

Initial relative quantity

I1 I2 I8

Re

lativ

e q

ua

ntity

by

LH-P

CR

0

2

4

6

8

10

Fig. S5 Evaluation of LH-PCR amplification efficiencies of miRNA and its precursor.

RNA

Pre-miR-122 miR-122Re

lativ

e a

mp

lific

atio

n e

ffe

cie

ncie

s

0

20

40

60

80

100

120

miR-200b family

miR-200a miR-200c miR-200bRe

lativ

e a

mp

lific

atio

n e

ffe

cie

ncie

s

0

20

40

60

80

100

120

Fig. S6 Evaluation of LH-PCR amplification efficiencies of miRNA and its family members.

miR-29a

miR

-29a

miR

-29b

miR

-29c

Tissue RNA

Fig. S7 Detection of miR-29 family members by conventional Northern blotting.

Rno-miR-138

pre Hela liver kidey lung brain

Fig. S8 Detection of miR-138 and its precursor from tissue samples by stem loop-PCR .