IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-12: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
|||
| Line 1: | Line 1: | ||
==Results of Restreak of J00450+PSB4A3== | |||
[[Image:Restreak 81206 zs.jpg]] | [[Image:Restreak 81206 zs.jpg]] | ||
Although the colonies took two days to grow, the restreak of the plates on Carb worked; we will now verify the ligation via PCR while growing liquid culture from them. | |||
===PCR ligation test of J04450+PSB4A3== | |||
Following Perry's Protocol which worked pretty well, in order to check the ligation we will do a PCR using the PCR Supermix from the VF2 and VR areas of the plasmid; | |||
From the above image, we will try 7 colonies: | |||
*Top10 plate2 A (eg. 102) | |||
*Top10 plate1 A (eg. 101) | |||
*Top10F plate1 (eg. F1) | |||
*Top10F plate2 1 (eg. F2-1) | |||
*Top10F plate2 2 (eg. F2-2) | |||
*Top10F plate2 3 (eg. F2-3) | |||
*Top10F plate2 3 (eg. F2-4) | |||
From Perry's Protocol, each will have: | |||
*8uL PCR supermix | |||
*1uL VF2 (2uM) | |||
*1uL VR (2uM) | |||
Then, get a carb plate, pick the desired colony, streak it on the plate, and then dip it into the 10uL rxn. | |||
Revision as of 12:53, 12 August 2006
Results of Restreak of J00450+PSB4A3
Although the colonies took two days to grow, the restreak of the plates on Carb worked; we will now verify the ligation via PCR while growing liquid culture from them.
=PCR ligation test of J04450+PSB4A3
Following Perry's Protocol which worked pretty well, in order to check the ligation we will do a PCR using the PCR Supermix from the VF2 and VR areas of the plasmid;
From the above image, we will try 7 colonies:
- Top10 plate2 A (eg. 102)
- Top10 plate1 A (eg. 101)
- Top10F plate1 (eg. F1)
- Top10F plate2 1 (eg. F2-1)
- Top10F plate2 2 (eg. F2-2)
- Top10F plate2 3 (eg. F2-3)
- Top10F plate2 3 (eg. F2-4)
From Perry's Protocol, each will have:
- 8uL PCR supermix
- 1uL VF2 (2uM)
- 1uL VR (2uM)
Then, get a carb plate, pick the desired colony, streak it on the plate, and then dip it into the 10uL rxn.
