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Edinburgh/15 August 2008
From 2008.igem.org
Week 9
Friday 15 August 08
- Plates from yesterdays transformations of pSB1A2+cex and pSB1A2+cenA (119-122) failed to show any signs of growth. The transformation is being repeated. (AH)
- Glycogen assay performed: Flooding colonies with Gram's iodine solution yields results after ~5 minutes. Colonies on the (+)glucose plate stained darker than those on the (-)glucose plate (brown compared to no apparent change). The staining seems to grow more apparent up to ~30 minutes and then fades to nothing as the plate dries off. (AH)
- Transformed with L39 (pSB1A2+cex), L40 (pSB1A2+cenA) and L41 (pSB1A2+dxs+crtE) onto Plates 123 to 128.(HX)
- Minipreps of pSB1A2+crtE from its corresponding cultures 1 to 4 made M139 to M143. (M142 and M143 made from the same culture (4). M143 has double quantity of solution 1.) Then double digests of M139 to M143 with EcoRI/PstI and the products were run on Gel 53. (Yan, OG)
- PCR of M43 (repeat: glgC-mut1,2, P71A), M120 (repeat: glgC-mut1,2,3, P72), rrnB from the four C. fimi genomic DNA preps made yesterday, plus the original prep as a positive control (P73-77). PCR products run on Gel 54. Results:
- P71A and P72 succeeded, producing large quantities of DNA around the length of glgC
- P74 yielded a single band around 1.5-2kb
- P73 and P75 yielded gel-length smears
- Nothing came of P76 and P77 (the control). (AM)
