IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-30: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
JeffreyLau (talk | contribs) |
JeffreyLau (talk | contribs) |
||
| Line 26: | Line 26: | ||
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). | 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. | We also inoculated 12 liquid cultures (2 mL LB Kan+Amp), one for each colony, from the restreak plates. | ||
Revision as of 20:58, 30 October 2006
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
J36337 (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.
