Edinburgh/20 August 2008

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(New page: === Week 10 === ==== Wednesday 20 August ==== *Transformation of L48 into plate 136/137 (dxs+Lims+AppY), and L49 into plate 138/139 (CrtBI+Appy) (YAN) Note: a control was set up by trans...)
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=== Week 10 ===
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<div id="header">{{Template:Team:Edinburgh/Templates/Header}}</div>
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==== Wednesday 20 August ====
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*Transformation of L48 into plate 136/137 (dxs+Lims+AppY),  and L49 into plate 138/139 (CrtBI+Appy) (YAN) Note: a control was set up by transforming only 1.5 microlitre of Edinbrick, as the newly made competent cells were used for transformation, made *Plate 140/141(*Edinbrick1 only).
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'''[[Edinburgh/19_August_2008|< Previous Entry]] | [[Team:Edinburgh/Notebook|Notebook Index]] | [[Edinburgh/21_August_2008|Next Entry >]]'''
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*Double digestion of Lac Z (Lablled as RBS+Lac Z in Edinbrick1 from iGEM 06) using Xbal/PstI, then ligated into PcstA, which has been digested and purified. (vector).  
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*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)  
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== Week 10 ==
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M156-M159: Minipreps of cenA, digested DNA and ran on Gel 58(OG)  
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=== Wednesday 20 August ===
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*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)  
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*Hence, P78~P83 were purified and self-ligated (L50~L55). (AM)  
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*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)
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*PCR of Zea mays genes from maxipreps: zm1 (ISA1) from X11 (P84), zm2 (ISA2) 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)  
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*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).
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*Analytical digests of X1-X17 with EcoR1-Pst1. X1-X13 run on Gel 57 and X14-X17 run on Gel 58. (HX)   
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*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)
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*Ligation of Lac Z into PcstA (vector) (L50) (yan)
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*M156-M159: Minipreps of cenA, digested DNA and ran on Gel 58(OG)  
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*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)  
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*Hence, P78~P83 were purified and self-ligated ('''L50~L55'''). (AM)  
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*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)  
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*Analytical digests of X1-X17 with EcoR1-Pst1. X1-X13 run on '''Gel 57''' and X14-X17 run on '''Gel 58'''. (HX)   
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*Ligation of ''lacZ'' into P''cstA'' (vector) ('''L50''') (yan)

Latest revision as of 21:53, 29 October 2008

< Previous Entry | Notebook Index | Next Entry >

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)
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