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Edinburgh/26 August 2008
From 2008.igem.org
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Week 11
Tuesday 26 August 08
- Sequencing results for 150 and M156 received. These show that the suffix for M150 is correct (and that it is preceded by cex), but M156 does not contain cenA. (AH)
- Submitted M160 (pSB1A2+dxs+lims+appY) for sequencing with primers pSB1A2insF2 and pSB1A2insR2. (AH)
- Gel 63: digests of M175-M181: (L52, SOB+glgC-mut1,2,3), M176-M178 are from plate 161 streaks 1-3 (L53, SOB+rbs+glgC-mut1,2) and M179-M181 are from plate 163 streaks 1-3 (L56, pSB1A2+PcstA+lacZ). (YAN)
- Re-digests of M160, M164-M174:
- M160: pSB1A2+dxs+lims+appy
- M164: plate 155 streak 5 (L49, pSB1A2+crtB+crtI+appY)
- M165-M168: plate 158 streaks 1-4 (L50, SOB+glgC-mut1,2)
- M169-M172: plate 159 streaks 1-4 (L51, SOB+glgC-mut1,2,3)
- M173: plate 160 streak 1 (L55 (SOB+rbs+glgC-mut1,2,3)
- M174: L52, SOB+glgC-mut1,2,3)
- Run on Gel 65. Results: M168-172 showed good results with SOB ~ 2.0kb, glgC ~1.3kb, incomplete digest ~3.3kb. (YAN)
- Also digested M166-168 (SOB+glgC-mut1,2) and M169-M171 (SOB+glgC-mut1,2,3) with a) EcoRI and b) EcoRI/SpeI as the results for these looked reasonable on gel 62. (AH)
- PCR of beta-glucosidase using cytophaga as template, anealing temp 55C and extension time of 30 seconds, P90. (YAN)
- PCR products of P88 (SU1) and P89 (ISO2) were run on Gel 64. The desired bands were present along with several others.(YAN)
- Gel 66: Repeat of Gel 64 (P68 and P79) with the addition of P90 (CHU2268 bglX). Stained with SYBRsafe, hence no photo. Desired bands from P88 and P89 were cut out for purification tomorrow. P90 revealed a single band in the desired location. (AM)
