Team:Hawaii/Test Competent E. Coli

From 2008.igem.org

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Next time, remember to aliquot out and transform only 50 ul of the competent cells.
Next time, remember to aliquot out and transform only 50 ul of the competent cells.
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* What was learned and how to do future experiments differently.
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<blockquote>''Insanity is doing the same thing over and over again and expecting different results.'' - Albert Einstein</blockquote>
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{{Team:Hawaii/Footer}}
{{Team:Hawaii/Footer}}

Revision as of 01:32, 4 June 2008

Contents

Test Competent E. coli DH5a

  • Test our competent cells against Callahan competent cells

Methods

Materials

  • Transported 50uL of competent cells from Callahan Lab on ice
  • 100uL batch 1 competent cells made at OD600 ~0.6 (split into two 50 uL aliquots 4-18a,4-18b)
  • 100uL batch 2 competent cells made at OD600 ~0.3 (split into two 50 uL aliquots 2-17a,2-17b)

Procedure

  1. Thaw plasmid on ice
  2. Add 1uL plasmid pBluescript
  3. Incubate plasmid+competent cells on ice for 30 minutes
  4. Heat shock 60 sec at 42C
  5. Add 250 μl SOC
  6. Incubate at 37 C for 1 hour in 2 ml eppendorf tubes, with shaking (150 rpm)
  7. Plate 20 uL dilutions 1/1, 1/10, 1/100 to test competency

Results


Discussion

Next time, remember to aliquot out and transform only 50 ul of the competent cells.


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