Waterloo/15 July 2008
From 2008.igem.org
08/07/15 Tues
[x] observe results from competent cells test, rfp, C0052', C0051', streaked placgfp to UV competent cells
- placgfp grew, [rfp, C0052',C0051' did not grow, will religate]
[x] expose streaked placgfp plates to UV
- didn't work
[] colony PCR of CI' (51) and CI' (52)
[] inoculation
[] diagnostic gel
[] test Pfu with new buffer at the same time
- no growth in transformants, so will religate and retransform today
[x] ligation (repeated) of C0052', C0051', rfp'
[x] transformation of ligation
[x] full digestion of C0052, C0051, rfp, TT, RBS
[x] gel of full digestion
[x] bands cut out of gel and put in the -20
[x] aliquot SpeI and ligase buffer
[x] full digest of c0051, TT cut and uncut
[x] gel of full digest
- excised probable band for TT (in -20) but digest for cI failed
[x] inoculate [TT, Pconst, RBS, rfp'.] from frozen stock, [C0052, R0010, R0040 B0034, C0051, C0052] from plates on bench.
[x] streak out POcI-rfp