Team:NTU-Singapore/Notebook/23 July 2008

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(New page: =Wednesday 23 July= Hung, Min: ==Digestion for Terminator with X/P on 2% gel== *As we could not see the band of Terminator insert on Tuesday to extract) *Gel loading and extraction + nanod...)
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=Wednesday 23 July=
=Wednesday 23 July=
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Hung, Min:
Hung, Min:
==Digestion for Terminator with X/P on 2% gel==
==Digestion for Terminator with X/P on 2% gel==
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*Lysis(I)-Empty standard vector: digested DNAs from -20 fridge.  
*Lysis(I)-Empty standard vector: digested DNAs from -20 fridge.  
*LsrA(I)-Empty standard vector: digested DNAs from -20 fridge.<br>'''NOTE:'''After ligation, we used QIAgen PCR purification kit to purify (instead of MinElute as the columns wouldn't arrive until next week). Therefore, when transforming into the cells, we used 9ul (from 30ul after purification) instead of 3ul.
*LsrA(I)-Empty standard vector: digested DNAs from -20 fridge.<br>'''NOTE:'''After ligation, we used QIAgen PCR purification kit to purify (instead of MinElute as the columns wouldn't arrive until next week). Therefore, when transforming into the cells, we used 9ul (from 30ul after purification) instead of 3ul.
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Revision as of 12:23, 25 October 2008


Contents

Wednesday 23 July

Hung, Min:

Digestion for Terminator with X/P on 2% gel

  • As we could not see the band of Terminator insert on Tuesday to extract)
  • Gel loading and extraction + nanodrops.
  • stored at -20 fridge for later ligation.

Minipreps

  • for the 12 samples inoculated on Tuesday (including Lysis-Term,Term,E7 and T7ptag)
  • Digestional gel check: we did normal digestion (E/P for 2 hours):
    • 3 lysis-term.
    • 1 Term (for comparision with Lysis-Term)
    • 1 Lysis (for comparision with Lysis-Term)
    • 1 E7
    • 1 T7ptag
  • We also tested Fast digestion for T7ptag for 5 mins.
    RESULT: As we used 1kb ladder, we actually could not tell whether the Lysis-Term bands were correctly ligated (about 250bp) or they're just Term(129bp). We'll check again on Thursday, using 2% gel and 100bp ladder.

Ligation and Transformation:

  • E7(V)-Term(I): E7(V) taken from -20 fridge (from Tuesday); Term (I) was obtained today.
  • Lysis(I)-Term(V): we just transformed another 3ul of ligation mixture among 7ul left from Monday into cells.
  • Lysis(I)-Empty standard vector: digested DNAs from -20 fridge.
  • LsrA(I)-Empty standard vector: digested DNAs from -20 fridge.
    NOTE:After ligation, we used QIAgen PCR purification kit to purify (instead of MinElute as the columns wouldn't arrive until next week). Therefore, when transforming into the cells, we used 9ul (from 30ul after purification) instead of 3ul.
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