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

Data distribution/ storage at the MPI Berlin

Richard Reinhardt, MPI Molecular Genetics Berlin-Dahlem

http://est.molgen.mpg.de/SleepingBeauty/

User: SB-GroupPW : aAh1Hi2F