Team:NTU-Singapore/Notebook/22 July 2008

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=Tuesday 22 July=
=Tuesday 22 July=
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{|border="1" style="background-color:#ffffcc;" cellpadding="20"
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Hung,Lu Chao, Min:
Hung,Lu Chao, Min:
==Ligation of E7 (vector) with Terminator (insert)==
==Ligation of E7 (vector) with Terminator (insert)==
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*1400: staining
*1400: staining
*1430: Gel extraction, nanodrops. After that, we stored the digested DNAs (ready for ligation) at -20 fridge for ligation on Wednesday.
*1430: Gel extraction, nanodrops. After that, we stored the digested DNAs (ready for ligation) at -20 fridge for ligation on Wednesday.
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==Ligation of lysis and LsrA into standard vector:
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==Ligation of lysis and LsrA into standard vector:==
*Cut Lysis,LsrA and GFP with E/P in Buffer 2, incubated for 2 hours.
*Cut Lysis,LsrA and GFP with E/P in Buffer 2, incubated for 2 hours.
*1345: QIA PCR purification
*1345: QIA PCR purification
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==Ligation of Lysis (I) and Terminator (V):==
==Ligation of Lysis (I) and Terminator (V):==
*Do the same as Monday, in case all 3 colonies of Lysis-Term. are wrong.
*Do the same as Monday, in case all 3 colonies of Lysis-Term. are wrong.
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Latest revision as of 02:32, 28 October 2008


Contents

Tuesday 22 July

Hung,Lu Chao, Min:

Ligation of E7 (vector) with Terminator (insert)

  • E7: cut with S/P for 2 hours
  • Terminator: cut with X/P for 2 hours
  • 1245: QIA PCR purification
  • 1300: gel loading (each sample into 3 lanes)
  • 1400: staining
  • 1430: Gel extraction, nanodrops. After that, we stored the digested DNAs (ready for ligation) at -20 fridge for ligation on Wednesday.

Ligation of lysis and LsrA into standard vector:

  • Cut Lysis,LsrA and GFP with E/P in Buffer 2, incubated for 2 hours.
  • 1345: QIA PCR purification
  • 1400: gel loading (each sample into 3 lanes)
  • 1500: staining
  • 1530: Gel extraction, nanodrops. After that, we stored the digested DNAs (ready for ligation) at -20 fridge for ligation on Wednesday.

Inoculation:

  • 3 lysis-terminator colonies transformed on Monday.
  • 3 E7-empty vector and 3 T7ptag-empt vector colonies.
  • 3 Terminator colonies (obtain more terminator plasmid for ligation).

Ligation of Lysis (I) and Terminator (V):

  • Do the same as Monday, in case all 3 colonies of Lysis-Term. are wrong.
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