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Edinburgh/20 August 2008
From 2008.igem.org
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=== Wednesday 20 August === | === Wednesday 20 August === | ||
| - | *Transformation of L48 into | + | *Transformation of L48 into '''Plates 136/137''' (''dxs+LIMS+appY''), L49 into '''Plates 138/139''' (''crtBI+appY''), and control with only 1.5µl Edinbrick1 into '''Plates 140/141'''. (YAN) |
| - | *Double digestion of | + | *Double digestion of ''lacZ'' (Lablled as RBS+''lacZ'' in Edinbrick1 from iGEM 06) using Xbal/PstI, then ligation into P''cstA'', 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) | + | *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) | 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) | + | *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) | + | *Hence, P78~P83 were purified and self-ligated ('''L50~L55'''). (AM) |
| - | *PCR of Zea mays genes from maxipreps: | + | *PCR of Zea mays genes from maxipreps: ''SU1'' from X11 ('''P84'''), ''ISO2'' 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) | + | *Analytical digests of X1-X17 with EcoR1-Pst1. X1-X13 run on '''Gel 57''' and X14-X17 run on '''Gel 58'''. (HX) |
| - | *Ligation of | + | *Ligation of ''lacZ'' into P''cstA'' (vector) ('''L50''') (yan) |
Revision as of 21:53, 29 October 2008
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Week 10
Wednesday 20 August
- Transformation of L48 into Plates 136/137 (dxs+LIMS+appY), L49 into Plates 138/139 (crtBI+appY), and control with only 1.5µl Edinbrick1 into Plates 140/141. (YAN)
- Double digestion of lacZ (Lablled as RBS+lacZ in Edinbrick1 from iGEM 06) using Xbal/PstI, then ligation 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: SU1 from X11 (P84), ISO2 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 lacZ into PcstA (vector) (L50) (yan)
