User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/15: Difference between revisions

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#* 750μL pTXB1His/DH10B QuikChange colony #1 culture from [[User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/14| yesterday]] + 250 μL 60% glycerol (sterile)
#* 750μL pTXB1His/DH10B QuikChange colony #1 culture from [[User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/14| yesterday]] + 250 μL 60% glycerol (sterile)
#*→ -80°C
#*→ -80°C
# [[Muratore:Protocols/DNA purification/Miniprep| Miniprep]] of pTXB1His
# [[Muratore:Protocols/DNA purification/Miniprep| Miniprep]] of pTXB1His and pTXB1
## 10mL LB + 10μL 100 mg/mL Amp + pTXB1His QC colony #2 (see [[#Results| Results below]])
## 10mL LB + 10μL 100 mg/mL Amp + pTXB1His QC colony #3 (see [[#Results| Results below]])
## 10mL LB + 10μL 100 mg/mL Amp + pTXB1 colony (from [[User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/09| last week]])
##*→ 37°C O/N w/ shaking


==Results==
==Results==

Revision as of 14:46, 15 June 2011

AU CHEM-570 Lab Prep <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Benchwork

Plan

  • Miniprep pTXB1His and send for sequencing
  • Start more cultures of new colonies for more minipreps
  • Don't forget to miniprep pTXB1 (unmodified) as well
  • Streak expression strain (ER2566) on LB so competent cells and a glycerol stock can be made

Experiments

  1. Glycerol stock
    • 750μL pTXB1His/DH10B QuikChange colony #1 culture from yesterday + 250 μL 60% glycerol (sterile)
    • → -80°C
  2. Miniprep of pTXB1His and pTXB1
    1. 10mL LB + 10μL 100 mg/mL Amp + pTXB1His QC colony #2 (see Results below)
    2. 10mL LB + 10μL 100 mg/mL Amp + pTXB1His QC colony #3 (see Results below)
    3. 10mL LB + 10μL 100 mg/mL Amp + pTXB1 colony (from last week)
      • → 37°C O/N w/ shaking

Results

  • Four plates yesterday:
    • 200μL experimental transformation from yesterday: 1 colony
      • → pTXB1His QC colony #2
    • 200μL control transformation from yesterday: 0
    • 800μL experimental transformation re-plated from 2 days ago: 1 colony
      • → pTXB1His QC colony #3
    • 800μL control transformation re-plated from 2 days ago: 0
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