Melbourne/Lab Notebook Weeks 5-8

From 2007.igem.org

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*[[Melbourne/Diagnostic Digest|Digested]] minipreprs from 9/7:
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*# [[Melbourne/BBa_E0430|P1 11A]] with X/P
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*# [[Melbourne/BBa_R0082|P1 15P]] with S/P
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(no BSA or AP used).
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*Inactivated enzymes at 80degreesC in a heat block for 10min.
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*[[Melbourne/Ligation Protocol|Ligated]] the two parts together (5ul each) overnight at 4degreesC.
<font size=3><b>31 July 2007  
<font size=3><b>31 July 2007  
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*[[Melbourne/Transformation Protocol|Transformation]] of ligations from 31/7 into NM522 competent cells using 3ul of DMSO.
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*[[Melbourne/Diagnostic Digest|Diagnostic Digest]] of  
*[[Melbourne/Diagnostic Digest|Diagnostic Digest]] of  
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*[[Melbourne/Growing up cells|Cultured]] 6 colonies picked from 1/8 transformations. A total of about 500 colonies grew with none on control (competent cells only plate).
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*[[Melbourne/Transformation Protocol|Transformation]] of [[Melbourne/BBa_E0033|P2 9E]] (LacZa with KanR) from iGEM plate.  
*[[Melbourne/Transformation Protocol|Transformation]] of [[Melbourne/BBa_E0033|P2 9E]] (LacZa with KanR) from iGEM plate.  
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*[[Alkaline Lysis Miniprep protocol|Alkaline Lysis Miniprepped]] colonies from 2/8. ([[Melbourne/BBa_E0430|P1 11A]] and  [[Melbourne/BBa_R0082|P1 15P]])
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-results: colony 6 had two bands of sizes approximately 1000 and 1100bp.
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*Streaked out colony 6 liquid culture on LBA plate and grew overnight.

Revision as of 13:15, 8 October 2007

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Contents

Week 5

23 July 2007

  • Colony PCRed using vf2 and vR primers the transformations from 20/7. 3 colonies from ligation plate and one from plasmid control plate.

24 July 2007

-results: colony 3 didn't give product.

  • Streaked out some cells from index plate (No.3).


25 July 2007


26 July 2007


27 July 2007

Week 6

30 July 2007

(no BSA or AP used).

  • Inactivated enzymes at 80degreesC in a heat block for 10min.
  • Ligated the two parts together (5ul each) overnight at 4degreesC.

31 July 2007

  • Transformation of ligations from 31/7 into NM522 competent cells using 3ul of DMSO.


using miniprep from 13/7. All digests were E/P.

Ran a Gel:

-Results:

    2. Low digestion efficiency
    3. Low digestion efficiency
    4. No insert seen but DNA present
    5. No insert seen but DNA present
    6. Ok digestion (not very good)
    7. Ok digestion (not very good)

--Digestion imcomplete??

1 Aug 2007

  • Cultured 6 colonies picked from 1/8 transformations. A total of about 500 colonies grew with none on control (competent cells only plate).


2 Aug 2007

-results: colony 6 had two bands of sizes approximately 1000 and 1100bp.

  • Streaked out colony 6 liquid culture on LBA plate and grew overnight.


3 Aug 2007


4 Aug 2007

Week 7

5 Aug 2007


6 Aug 2007


7 Aug 2007


8 Aug 2007


9 Aug 2007


10 Aug 2007


11 Aug 2007

Week 8

12 Aug 2007


13 Aug 2007


14 Aug 2007


15 Aug 2007


16 Aug 2007


17 Aug 2007


18 Aug 2007

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