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Edinburgh/Notebook/PCR products
From 2008.igem.org
P1: dxs from Escherichia coli JM109. (25.06.08)
P2: appY from E. coli JM109. (25.06.08)
P3: glgC from E. coli JM109. (25.06.08)
P4: fusion PCR for BABEL1+glgC. (01.07.08)
P5: fusion PCR for BABEL2+glgC. (01.07.08)
P6: mutagenic PCR to remove EcoRI site 1 from glgC (M11). (04.07.08)
P7: mutagenic PCR to remove EcoRI site 2 from glgC (M11). (04.07.08)
P8: crtB (from Pantoea ananatis cells). (08.07.08)
P9: crtI (from Douglas's maxiprep of the mutated crtIB plasmid). (08.07.08)
P10: crtY (from P. ananatis cells). Failed. (08.07.08)
P11: fusion PCR for rbs+dxs. Failed. (10.07.08)
P12: repeat fusion PCR for rbs+dxs, using L11 and primers rbs2fclon1+pSB1A2insr1. (13.07.08)
P13 and P14: MABEL mutation of site 1 on glgC mutant clones M19 and M22.
P15: fusion PCR for rbs+appY
P16, P17, P18 and P19: first attempts at cenA, cenB, cenC and cex.
P20, P21, P22 and P23: second attempts at cenA, cenB, cenC and cex.
P24, P25: repeat PCR for cenA and cex using Pfu. Failed.
P26: Positive control PCR with primers fd1 and rd1 to amplify 16S rRNA gene rrnB from Cellulomonas fimi.
P27: crtY from P. ananatis cells. - Failed
P28, P29: crtB and crtI from L18 and L19.
P30: Repeat positive control rrnB from C. fimi, annealing 1 minute.
P31: pZntA promoter
P32-P35: repeat PCR for cenA, cenB, cenC and cex, annealing 1 minute. Failed.
P36: M43 (glgC-mut1,2) with primers glgCf2/glgCr2 (25.07.08: AM)
P37: crtY from P. ananatis. (25.07.08: AM) - Failed
P38~P41: cenA, cenB, cenC and cex from heat-killed cell suspension.
P42~P45: cenA, cenB, cenC and cex from 'impure' DNA solution.
P46~P47: cenA and cex from 'impure' DNA solution, annealing 65C. Failed.
P48: crtY (new primer mixture) (previously P38a) (30.07.08: CF) - Failed
P49: Recreation of P12 (rbs+dxs) (previously P39a) (30.07.08: CF)
P50: third mutagenesis of glgc-mut1,2 (04.08.08: YAN)
P51: pCstA from E. coli cells (06.08.08: CF) - Failed
P52: cenA from heat killed C. fimi (07/08/08: YAN, AM) - Failed
P53: cex from heat killed C. fimi (07/08/08: YAN, AM) - Failed
P54: pCstA from E. coli cells - Purified (07.08.08: CF)
