Team:UNIPV-Pavia/Notebook/Week4Jul

From 2009.igem.org

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(July, 21st)
(July, 21st)
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**B0030(X3)
**B0030(X3)
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*We noticed unwanted bands in MRGENE1 run...So, we performed an analysis on Mr Gene plasmids and we found an unwanted PstI site in pMK-RQ that gives  
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*We noticed unwanted bands in MRGENE1 run...So, we performed an analysis on Mr Gene plasmids and we found an unwanted PstI site in pMK-RQ that gives the noticed bands...
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Revision as of 10:07, 22 July 2009

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December 2008
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March 2009
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April 2009
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May 2009
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June 2009
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July 2009
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August 2009
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September 2009
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October 2009
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November 2009
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Week from July 20th, to July 26th, 2009

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July, 20th

  • Overnight digestion (20 ul reaction volume) for:
MRGENE1(X-P) MRGENE1-2(X-P) MRGENE1-3(X-P)
MRGENE2(X-P)(X2) MRGENE2-2(X-P) MRGENE2-3(X-P)
B0030(S-P)(X3)
  • NOTE: reactions which involve excision(X-P) were performed on 3 ug of DNA, while S-P digestions were performed with 1 ug of DNA. We decided to have 3 replicates to be sure to have an acceptable yield.
  • We decided to perform gel run/cut/purification for MRGENE2 and for B0030, while we decided to perform DNA precipitation with sodium acetate for MRGENE1.



  • We transformed about 2.5 pg of A14-4 in TOP10 in order to filter the wanted plasmids. We plated the transformed bacteria and incubated the plates at 37°C overnight.



  • We infected 5 ml of LB + Amp with 10 ul of A12-2 and A12-3 glycerol stocks. Tomorrow they will be miniprepped and sent to sequence!



Preparation of experiment with Tecan F200

  • We infected 5 ml of LB + Kan with a single colony taken from the native plate of B0015.
  • We incubated the inoculum for 6 hours (37°C, 220 rpm).



Experiment with Tecan F200

  • Description
  • Purpose:
  • Materials & Methods
  • Protocol
  • Results


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July, 21st

  • Gel run for:
    • MRGENE1(for check)
    • MRGENE2
    • MRGENE2-2
    • MRGENE2-3
    • B0030(X3)
  • We noticed unwanted bands in MRGENE1 run...So, we performed an analysis on Mr Gene plasmids and we found an unwanted PstI site in pMK-RQ that gives the noticed bands...

Top

July, 22nd

Top


July, 23rd

Top


July, 24th

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