Uppsala-Sweden/6 September 2009

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Revision as of 23:09, 5 September 2009

Moving ON to fride

ON cultures for the lux system were moved to the iGEM fridge for MiniPrep tomorrow

Checking Plates

All plates show colonoies. After picking colonies -> fridge. --Karl.brune 23:08, 5 September 2009 (UTC)

Picking Colonies

I picked colonies from plates into 10µl selective media. --Karl.brune 23:08, 5 September 2009 (UTC) 5 Colonies of pirA (AC3) -> pirA-X 5 Colonies of b0034-pirA (AC3) -> RA-X 5 Colonies of B0034-Yadh2-B0015 (AC3) -> RYB-X 5 Colonies of B0015 (AC3) - T-X 5 Colonies of R0011-B0034 (AK3) -> RR-X

3 Colonies of E1A1-B0015 (AK3) -> E1A1X 3 Colonies of E1A2-B0015 (AK3) -> E1A2X 3 Colonies of E1A3-B0015 (AK3) -> E1A3X 3 Colonies of E1A4-B0015 (AK3) -> E1A4X

3 Colonies of E1B1-B0015 (AK3) -> E1B1X 3 Colonies of E1B2-B0015 (AK3) -> E1B2X 3 Colonies of E1B3-B0015 (AK3) -> E1B3X 3 Colonies of E1B4-B0015 (AK3) -> E1B4X

3 Colonies of E21-B0015 (AK3) -> E21X 3 Colonies of E22-B0015 (AK3) -> E22X 3 Colonies of E23-B0015 (AK3) -> E23X 3 Colonies of E24-B0015 (AK3) -> E24X

X= # of colony

--Karl.brune 23:08, 5 September 2009 (UTC)

Colony PCR

TAQ colony PCR was performed.

10µl reaction, standart mix with verfication primer VF2 and VR.

Initial Denat: 1min 30sec @94°C

40Cylces

Denaturation: 45 sec @94°C

Annealing: 45 sec @50°C

Elongation: 1min 45sec @ 72°C

Return to Denat.

Final Elongation: 10min @72°C

Infinite storage at 4°C

Colony tubes were moved to fridge after taking template for the PCR