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Edinburgh/21 August 2008
From 2008.igem.org
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Week 10
Thursday 21 August
- Submitted M156 (pSB1A2+cenA) for sequencing with primer pSB1A2insF2. (AH)
- Performed single (PstI) and double (EcoRI/PstI) digests of X2 (pSB1A2+dxs), X9 (pSB1A2+rbs+dxs), X16 (pSB1A2+PcstA) and X17 (pSB1A2+dxs+lims). These were run on gel 59. Results look good for X2, X9 and X16, but X17 appears to be wrong. (We must have maxiprepped the wrong colony, so maxipreps will need to be performed for the other colonies, followed by analytical digest.) (AH)
- Subplated some colonies from yesterday's transformations of L48 and L49. There were only a few colonies per plate, and worryingly none of them are blue. However, there were loads of blue colonies on the Edinbrick1 control plates implying that there isn't anything wrong with the competency of the cell. Although there were so few colonies, emergence of satellite colonies necessitated the subplating onto plate 155:
One colony from Plate 137 (L48)
Four colonies from plate 138 (L48)
One colony from plate 139 (L49)
There was no growth on plate 140 (L49), so this, along with Plates 137 and 139, have been left at 37C to see if there is any more growth. Plate 138 has been stored at 4C. (AH)
- Maxprep culture of Plate 115, streak 2, 3 & 5.(HX)
- Transformation of L50~L56 into Plates 142~154. (AM)
- PCR of Zea mays genes from maxipreps: SU1 from X11 (P86), ISO2 from X12 (P87). Standard PCR conditions for KOD, annealing 62°C, extension 75s. Run on Gel 60. Results: even worse than yesterday's. P86 gave a long smear with no distinguishable bands. (AM)
