IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-30

Making stabs

 * Heat up LB agar 10min @ 50% power in microwave
 * Aliquot out 20mL
 * Add 40uL 50mg/uL AMP
 * Aliquot 1mL into each stab

Stabs made
BBa_J  name           frozen stock 36801  KaiA           2006_8_13KaiA 36802  KaiB           2006_8_13KaiB 36804  KaiC           2006_8_13KaiC 36831  Lac+RBS+A      A9 green tube 36832  Lac+RBS+B      colony E 10.21 36834  Lac+RBS+C      M2-7 8.30 36335  LacRBSALacRBSC colony 1 10.21 36336  LacRBSBLacRBSC J36021 10/26/06

Updated biobricks registry & sent parts
yup

J36937 (A+B+C) transformants
We have 12 colonies, labeled A-L.

Restreaked A-L on two LB Kan plates.

Performed diagnostic PCR (8 uL PCR Supermix, 1 uL 2nM VF2, 1 uL 2nM VR primers) on each of the 12 colonies, along with a positive control (J36831 colony) and negative (no colony).

PCR schedule:
 * 95C for 15'00
 * Do 30 times:
 * 95C for 0'30
 * 55C for 0'30
 * 72C for 3'30
 * 72C for 10'00
 * 4C forever

PCR will finish around 0140.

We also inoculated 12 liquid cultures (2 mL LB Kan+Amp), one for each colony, from the restreak plates.

Inoculation of J36834 (C), J36335 (A+C), J36336 (B+C)
Inoculated 2 mL LB Kan+Amp of J36834 (C), J36335 (A+C), J36336 (B+C).