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
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