Edinburgh/14 August 2008


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Thursday 14 August 08

 * Subplated from plate 116 (pSB1A2+crtE 100μl) onto plate 118. Should be ready to make culture for miniprep later this afternoon. (AH)
 * Submitted M121 (pSB1A2+crtY), M124 (pSB1A2+rbs+crtY), M130 (pSB1A2+PcstA) and M137 (pSB1A2+dxs+lims) for sequencing using pSB1A2insF2 for sequencing of all 4 plus pSB1A2insR2 for M137. Also resubmitted M109 (pSB1A2+crtB+crtI) for sequencing using both forward and reverse primers, because I realised that yesterday I circled BigDye rather than "reaction required" on the sequencing form! (AH)
 * Made culture of pSB1A2+crtE from plate 118, which was subcloned from plate 116 for minipreps (1-4) (Yan, HX).
 * Transformation of L40 (pSB1A2+cenA) onto plates 119/120 and L39 (pSB1A2+cex) onto plates 121/122. (Yan)
 * Digested and ligated M63 (pSB1A2+rbs+crtE) to M72 (pSB1A2+rbs+dxs). For M63 I used buffer H, XbaI/PstI. For M72 I used buffer B, SpeI/PstI. rbs+crtE inserted downstream of pSB1A2+rbs+dxs. The rbs+crtE and pSB1A2+rbs+dxs'' DNA was taken from a gel after SYBR-Safe staining. (OG)
 * Purified four preps of C. fimi genomic DNA from bottled cultures made by Dr. French (in LB, made from C. fimi nutrient agar plates on lab bench). (AM)
 * PCR of M43 (BABEL2+glgC-mut1,2, P70) and M120 (BABEL2+glgC-mut1,2,3, P71) with blunt-ended glgC primers. Run on Gel 52. Results: P70 failed; P71 yielded a thick band around 1.5kb (proper size for glgC) and two unexpected bands around 6kb and 12kb. (AM)


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