Registry/Measurement kit/Notebook/2007-6-29: Difference between revisions
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(New page: ==To do== #Colony PCR of promoter tester (We got 3 colonies) #Re-try colony PCR of RBS tester #Run them on a gel #MfeI/NsiI digest of E0240 #PCR cleanup E0240 #Ligate E0240 and 3K3, transf...) |
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*Trying extension time of 1:45, in case 1:30 was too short | *Trying extension time of 1:45, in case 1:30 was too short | ||
*Made new dNTP dilution (108 H2O + 3 of each NTP) | *Made new dNTP dilution (108 H2O + 3 of each NTP) | ||
==Analytic Gel== | |||
*Loading bottom row: | |||
*Lanes 1-5 and 16-20 empty or ladder; lanes 6-12 rbs tester; lanes 13-15 promoter tester. | |||
==Digest== | ==Digest== | ||
*Can't digest with both enzymes concurrently according to NEB, so cutting with MfeI first | *Can't digest with both enzymes concurrently according to NEB, so cutting with MfeI first | ||
Revision as of 12:39, 29 June 2007
To do
- Colony PCR of promoter tester (We got 3 colonies)
- Re-try colony PCR of RBS tester
- Run them on a gel
- MfeI/NsiI digest of E0240
- PCR cleanup E0240
- Ligate E0240 and 3K3, transform
Colony PCR
- Trying extension time of 1:45, in case 1:30 was too short
- Made new dNTP dilution (108 H2O + 3 of each NTP)
Analytic Gel
- Loading bottom row:
- Lanes 1-5 and 16-20 empty or ladder; lanes 6-12 rbs tester; lanes 13-15 promoter tester.
Digest
- Can't digest with both enzymes concurrently according to NEB, so cutting with MfeI first
