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Edinburgh/20 August 2008
From 2008.igem.org
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Week 10
Wednesday 20 August
- Transformation of L48 into plate 136/137 (dxs+Lims+AppY), and L49 into plate 138/139 (CrtBI+Appy) (YAN) Note: a control was set up by transforming only 1.5 microlitre of Edinbrick, as the newly made competent cells were used for transformation, made *Plate 140/141(*Edinbrick1 only).
- Double digestion of Lac Z (Lablled as RBS+Lac Z in Edinbrick1 from iGEM 06) using Xbal/PstI, then ligated into PcstA, which has been digested and purified. (vector).
- M150 and M151 (pSB1A2+cex) submitted for sequencing with primer pSB1A2insF2 as AH150F and AH151F. M152 (pSB1A2+dxs+crtE) submitted for sequencing with primers pSB1A2insF2 and pSB1A2insR2 as AH152F and AH152R. (AH)
M156-M159: Minipreps of cenA, digested DNA and ran on Gel 58(OG)
- Results of yesterday's PCR: When run on Gel 56, P78~P80 produced smears but also distinguishable bands for glgC, SOB2 and SOB2+glgC; P81~P83 produced bands for SOB2 and SOB2+glgC. (AM)
- Hence, P78~P83 were purified and self-ligated (L50~L55). (AM)
- PCR of Zea mays genes from maxipreps: zm1 (ISA1) from X11 (P84), zm2 (ISA2) from X12 (P85). Standard PCR conditions for KOD, annealing 60°C, extension 75s. Run on Gel 58. Results: too many bands to be of practical value. (AM)
- Analytical digests of X1-X17 with EcoR1-Pst1. X1-X13 run on Gel 57 and X14-X17 run on Gel 58. (HX)
- Ligation of Lac Z into PcstA (vector) (L50) (yan)
