User:Mbennie/Notebook/Lab Notebook/Notebook/2007/08/14
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Cellular Adhesion
- Gel
- Ran 1.5% gel for 30 minutes at 100V with samples (20ul sample with 5ul of loading dye)
- There is no band at the correct length
- Problem might be caused by PCR: primer is now binding to entire piece of DNA instead of just half of it
- PCR
- Template: 20ul PCR Supermix, .2ul VF2 and VR, .5ul miniprep DNA
- SS+Fos #1
- SS+Fos #2
- SS+Fos #3
- SS+JunB #1
- SS+JunB #2
- SS+JunB #3
- Protocol: Same as 8.1.2007
- Template: 20ul PCR Supermix, .2ul VF2 and VR, .5ul miniprep DNA
- Gradient PCR
- Template: 10ul PCR Supermix, .2ul BB_f and BB_r, .5ul ligation DNA (B+C with D)
- Ran eight samples from 55-70C on gradient PCR (other than annealing temp, everything else is the same as 8.1.2007 protocol)
- Template: 10ul PCR Supermix, .1ul IgAb-F and IgAb-R, .5ul ligation DNA (B+C with D)
- Ran eight samples from 55-70C on gradient PCR (other than annealing temp, everything else is the same as 8.1.2007 protocol)
- Template: 10ul PCR Supermix, .2ul BB_f and BB_r, .5ul ligation DNA (B+C with D)
- Gel
- Ran 1.5% gel for 30 minutes at 100V with samples (10ul sample with 4ul of loading dye)
- Looks like everything worked except SS+JunB #2 (~300bp)
- Gel
- Ran 1.5% gel for 30 minutes at 100V with samples (10ul sample with 4ul of loading dye)
- All bands are at 300bp
- WHAT IS THIS?!?!
- Possible Solution: By not PCR purifying tube D, it amplified D instead of the entire fragment
- PCR
- Template: 40ul PCR Supermix, .4ul VF2 and VR, .8ul DNA
- PCRed tube D to make some more
- Protocol: Same as 8.1.2007
- Template: 40ul PCR Supermix, .4ul VF2 and VR, .8ul DNA