High throughput technology group (htpt) R. Reinhardt (Max Planck Institute for Molecular Genetics,...
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Transcript of High throughput technology group (htpt) R. Reinhardt (Max Planck Institute for Molecular Genetics,...
High throughput technology
group (htpt)
R. Reinhardt
(Max Planck Institute for Molecular Genetics, Berlin)
Project overview: Joan Cerda
Richard Reinhardt, MPI Molecular Genetics Berlin-Dahlem
Joan Cerda Fundulus heteroclitus
(IRTA, Barcelona)
-1 cDNA synthesis,
Stage2 pool and normalize
-2
Stage3
-3
2 µg-10b- 3 (1)
10.
-10c-
-10a- Killifish 3 (1) 2 µg
3 (1) 2 µg
Project overview: Joan Cerda
Richard Reinhardt, MPI Molecular Genetics Berlin-Dahlem
Label Organism Stages Template check
cDNA normalize Size selection / ligation …
Picking Sequencing
SB10 Fundulus heteroclitus
(Killifish)
1,2,3 √ √ √ √ √ √ √ √ √ started (expected:
15.000 reads)
Project overview: Melody Clark
Richard Reinhardt, MPI Molecular Genetics Berlin-Dahlem
Label Organism Stage Template check, cDNA
normalize Size selection / ligation …
Picking Sequencing
SB5 Onychiurus arcticus
Control (untreated springtails cultured at +4 C)
√ √ √ √ started
SB6 Desiccated (taken from culture at –14 C)
√ √ √ √ started
SB7 Recovering(desiccated at –14 C; left to recover for 8 h with moisture)
√ √ √ √ started
SB8 Fully recovered (desiccated to –14 C; left to recover 24h)
√ √ √ √ started
SB9 Desiccating (taken to –2 C all starting the desiccation process)
√ √ in process - -
Project overview: Ora Hadas
Richard Reinhardt, MPI Molecular Genetics Berlin-Dahlem
Material for experiments to try the generation of bacterial cDNA libraries:
Aphanizomenon ovalisporum culture (BG=N; with heterocysts) [3] not started
Aphanizomenon ovalisporum culture (with akinetes) [4] not started
Fosmid libraries:
Label Sample DNA isolation
Size selection / ligation …
Picking Sequencing
SBFOS01 Aphanizomenon ovalisporum culture grown in full BG11 medium (i.e. no
akinetes, no heterocysts) [1]
√ √ completed (34x MTPs,
13056 clones)
in process (1416 reads)
SBFOS02 Aphanizomenon ovalisporum culture grown in full BG11 medium (with
akinetes, Kaplan) [2]
√ √ completed (40x MTPs,
15360 clones )
in process (2065 reads)
Project overview: Esther Lubzens
Richard Reinhardt, MPI Molecular Genetics Berlin-Dahlem
cDNA libraries:
Label Organism Stage Template check
cDNA normalize Size selektion / ligation …
Picking Sequencing
SB101 Rotifers POOL 1 √ (OD) √ in process - - -
SB102 POOL 2 √ (OD) √ in process - - -
SB103 POOL 3 √ (OD) √ in process - - -
SB106 see SB102 see SB102 see SB102 non-normalized - - -
Configuration of pools (alw ays taking in account the amounts of total RNA):
Mix1: mictic+amictic from clone C2 [1]/ mictic+amictic from clone [2] (1:1)
Mix2: mictic+amictic from clone1B4 [3]/ mictic+amictic from clone2A5 [4] (1:1)
Mix3: mictic+amictic from mixed population [5]
Pool: Mix1:Mix2:Mix3 (1:1:1)
POOL 2 Pool: RE dormant stage [8]/ RE dormant stage [9] (1:1)
POOL 3 Pool: RE 20 hr after initiating hatching [12]/ RE 20 hr after initiating hatching [14] (1:1)
POOL 1
Probably follow ing:
[SB104] subtraction SB101- mictic+amictic from clone1B4 [3]
[SB105] subtraction SB101- females bearing RE from clone 1B4 [6]
WAIT FOR RESULTS OF SB101/102/103