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Edinburgh/14 August 2008
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* 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) | * 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) | * 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 | + | * 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) |
Revision as of 11:49, 20 August 2008
Week 9
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)
