Edinburgh/Notebook/Minipreps

From 2008.igem.org

Revision as of 20:00, 6 October 2008 by Adlerma (Talk | contribs)

M1 to M6: pSB1A2+dxs transformants (plate 11). M2, M3, M4 and M6 all looked correct on gel 2. (30.06.08) - M2 was sequenced to confirm identity and was maxiprepped as X2.
M7: BABEL1+glgC transformant (plate 12). Did not look right on gel. (30.06.08)
M8 to M12: BABEL2+glgC transformants (plate 13). M10 and M11 both looked good on a gel, but sequencing showed that both had the insert in the reverse orientation. (30.06.08)
M13 to M18: pSB1A2+appY transformants. Gel 10 showed that M15 to M18, and possibly M14, all seemed to have inserts the right size. (09.07.08)
M19 to M24: BABEL2+glgC clones after mutation of EcoRI site 2. EcoRI digests on *Gel 11* suggest that *M19, M21 and M22* may have lost the EcoRI site. (10.07.08)
M25 to M30: possible rbs+dxs transformants from plate 34. (13.07.08)
M31 to M36: pSB1A2+crtB clones from plate 35. (14.07.08)
M37 to M42: pSB1A2+crtI clones from plate 36. (14.07.08)
M43 to M47: possible BABEL2+glgC double mutants (both EcoRI sites gone). (19.07.08)
M48: a possible pSB1A2+rbs+dxs clone. (19.07.08)
M49 to M54: pSB1A2-rbs+crtI. (29.07.08: CF)
M55 to M60: Putative pSB1A2+pZntA (L23). (29.07.08: Yan, Nimisha) - M50 sequenced (31.07.08: CF)
M61 to M66: Putative pSB1A2+rbs+crtE (L22). (29.07.08: Yan, Nimisha) - M63 sequenced (31.07.08: CF)
M67: rbs+crtB (L21) (29.07.08: Yan, Nimisha) - Sequenced (31.07.08: CF)
M68 to M71: pSB1A2+glgC-mut1,2 from plate 73 (31.07.08: AM, Yan)
M72 to M75: pSB1A2+rbs+dxs from plate 74 (31.07.08: AM, Yan)
M76 to M79: BABEL2+crtE from plate 74 (31.07.08: AM, Yan)
M80 to M83: pSB1A2+rbs+appY from plate 83. (04.08.08: Yan, OG)
M84 to M85: pSB1A2+rbs+crtB from plate 84. (04.08.08: Yan, OG)
M86 to M90: pSB1A2+cenA from plate 85. (04.08.08: Yan, OG)
M91 to M96: pSB1A2+cex from plate 86. (04.08.08: Yan, OG)
M97 to M104: pSB1A2+cenA from plate 85. (05.08.08: Yan)
M105 to M108: pSB1A2+cex from plate 85. (05.08.08: Yan)
M109 to M112: pSB1A2+crtB+crtI and plate 102 (11.08.08: OG)
M113 to M116: pSB1A2+glgC-mut1,2 and plate 103 (11.08.08: OG)
M117 to M120: pSB1A2+glgC-mut1,2,3 and plate 94 (11.08.08: OG)
M121 to M123: pSB1A2-crtY (12.08.08: OG)
M124 to M126: pSBA12-rbs+crtY (12.08.08: OG)
M127 to M130: pSB1A2-PcstA from plate 115 (13.08.08: OG)
M131 to M134: Minipreps of dxs+crtE (1,2,3,4) Note: M132 might be M136 (13.08.08: OG)
M135 to M138: Minipreps of dxs+lims (1,2,3,4) Note: M136 might be M132 (13.08.08: OG)
M139 to M143: Minipreps of pSB1A2 +CrtE (1 to 4) Note: M142 and M143 both made from culture 4.(15.08.08: YAN/OG)
M144 to M147: pSB1A2+cenA from plate 129 - Methanol used in preparation rather than ethanol! (18.08.08: OG)
M148 to M151: pSB1A2+cex from plate 129. (18.08.08: OG)
M152 to M155: pSB1A2+dxs+crtE from plate 130 (18.08.08: OG)
M156 to M159: pSB1A2+cenA (20.08.08: OG) M160 to M163: Plate 155 streaks 1-4 (L48, pSB1A2+dxs+lims+appY) (25.08.08: AH) M164: Plate 155 streak 5 (L49, pSB1A2+crtB+crtI+appY) (25.08.08: AH) M165 to M168: Plate 158 streaks 1-4 (L50, SOB+glgC-mut1,2) (25.08.08: AH) M169 to M172: Plate 159 streaks 1-4 (L51, SOB+glgC-mut1,2,3) (25.08.08: AH, Yan) M173: Plate 160 streak 1 (L55 (SOB+rbs+glgC-mut1,2,3) (25.08.08: Yan) M174 to M175: Plate 160 streaks 2 and 3 (L52, SOB+glgC-mut1,2,3) (25.08.08: Yan) M176 to M178: Plate 161 streaks 1-3 (L53, SOB+rbs+glgC-mut1,2) (25.08.08: Yan) M179 to M181: Plate 163 streaks 1-3 (L56, pSB1A2+pCstA+lacZ') (25.08.08: Yan) M182 to M187: pSB1A2+CenA (01.09.08: YAN) M188 to M193: CrtBI+AppY (01.09.08: YAN) M194 to M197: AppY+dxs+Lims (02.09.08:YAN) M198 to M200: L58 = self-ligation of P91 (SOB2-zm1). M199 and M200 were missed out by accident. (09.09.08: Yan) M201 to M203: L59 = self-ligation of P92 (SOB2-rbs-zm1). (09.09.08: Yan) M204 to M206: L60 = self-ligation of P93 (SOB2-zm2). (09.09.08: Yan) M207 to M209: L61 = self-ligation of P94 (SOB2-rbs-zm2). (09.09.08: Yan) M210 to M212: pSB1A2-glgc triple mutants. M213: dxs+CrtEBI. (15.09.08: AM) M214: pSB1A2-CenA. (15.09.08: AM) M215 to M216: pSB1A2-bglX. (15.09.08: AM) M217-M221: dxs+CrtEBI. (15.09.08: AM)