"
Team:The University of Alberta/7 August 2008
From 2008.igem.org
(Difference between revisions)
| Line 3: | Line 3: | ||
**Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands. | **Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands. | ||
***Ligated the gel purified product with J61003. | ***Ligated the gel purified product with J61003. | ||
| + | |||
| + | '''Chris''' | ||
| + | *PCR'd Tryp out of pUC57 (the original that Mike supplied)...buuut I used Vf and Vr to do it. I shouldnt have gotten any results because pUC57 doesnt have sites for those primers to bind BUT I did get a band about the right size...nevertheless, whatever was amplified wouldnt have the biobrick suffix and prefix in it so it was discarded. | ||
| + | *Redid the PCR with the correct primers....got a good, sharp band ~1500bp in size BUT the same band was in the negative control. I didnt want to risk using something that was just a random 1500bp fragment so.. | ||
| + | *Gel extracted the 1500bp fragment from the PCR sample | ||
| + | *I began sequencing it (Sequencing was finished by David and Kelly). Will take samples to MBSU tomorrow | ||
Revision as of 16:43, 8 August 2008
Saima
- PCRed PCAM2201 using Cam35s Forward and Reverse Primers. (Protocol to be followed is given in the Masterbook)
- Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands.
- Ligated the gel purified product with J61003.
- Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands.
Chris
- PCR'd Tryp out of pUC57 (the original that Mike supplied)...buuut I used Vf and Vr to do it. I shouldnt have gotten any results because pUC57 doesnt have sites for those primers to bind BUT I did get a band about the right size...nevertheless, whatever was amplified wouldnt have the biobrick suffix and prefix in it so it was discarded.
- Redid the PCR with the correct primers....got a good, sharp band ~1500bp in size BUT the same band was in the negative control. I didnt want to risk using something that was just a random 1500bp fragment so..
- Gel extracted the 1500bp fragment from the PCR sample
- I began sequencing it (Sequencing was finished by David and Kelly). Will take samples to MBSU tomorrow
