User:Eric Ma/Notebook/iGEM 2010 Lab Prep/2010/05/11

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(Test of Old Antibiotic Stocks)
(Test of Old Antibiotic Stocks)
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* This is a good sign. Antibiotic batches are still working.
* This is a good sign. Antibiotic batches are still working.
 +
==Test of RFP-expression constructs==
 +
* Found old P<sub>con</sub>-RBS-RFP-Terminator construct from the Lagally Lab -80ºC freezer. And gave a diabolical laugh when I finally found it after digging through the two -80ºC freezers we used for a whole 30 minutes.
 +
* Inoculated in Amp100 +/- arabinose.
 +
* Control: No amp, no arabinose, expect growth; next time need a negative control.
 +
* Setup as follows:
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{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Reagent'''
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| align="center" style="background:#f0f0f0;"|'''Tube A'''
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| align="center" style="background:#f0f0f0;"|'''Tube B'''
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| align="center" style="background:#f0f0f0;"|'''Tube C'''
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|-
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| Arabinose||+||-||-
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|-
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| Amp100||+||+||-
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|}
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* Results:
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Tube'''
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| align="center" style="background:#f0f0f0;"|'''Growth?'''
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| align="center" style="background:#f0f0f0;"|'''Red Coloration?'''
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|-
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| A||Yes||No
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|-
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| B||Yes||No
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|-
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| C||Yes||No
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|}
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* No red coloration observed when expected. Therefore, I will let the cultures grow a bit more to see if red coloration appears or not.
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Revision as of 14:00, 12 May 2010

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Test of old BW27783 + PBAD-RBS-RFP-Term Cultures

  • Found an old plate of BW27783s containing PBAD-RBS-RFP-Terminator in the 4ºC walk-in room
  • Tested by streaking on an Amp-100 plate + 0.1% (w/v) arabinose. Control: No arabinose
  • No growth observed on the next day.

Test of Old Antibiotic Stocks

  • Tested Amp100, Kan50 and Chlor25 from the old batch of antibiotics in the -20ºC freezer.
  • 3 ml LB + 3 µl antibiotics (each)
  • Inoculated using DH5α competent cells.
  • Results:
Amp100 Kan50 Chlor25 LB-only (Control)
No growthNo growthNo growthGrowth
  • This is a good sign. Antibiotic batches are still working.

Test of RFP-expression constructs

  • Found old Pcon-RBS-RFP-Terminator construct from the Lagally Lab -80ºC freezer. And gave a diabolical laugh when I finally found it after digging through the two -80ºC freezers we used for a whole 30 minutes.
  • Inoculated in Amp100 +/- arabinose.
  • Control: No amp, no arabinose, expect growth; next time need a negative control.
  • Setup as follows:
Reagent Tube A Tube B Tube C
Arabinose+--
Amp100++-
  • Results:
Tube Growth? Red Coloration?
AYesNo
BYesNo
CYesNo
  • No red coloration observed when expected. Therefore, I will let the cultures grow a bit more to see if red coloration appears or not.


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