Haynes Lab:Notebook/Short Projects/2014/09/21

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09/21/14 - Meli'sa Crawford

  • Restriction Digest of Plasmid DNA



Restriction Digest

1. 60ml of 1x-TAE

2. 6 grams Argarose (Low EEO)

3. Restriction Enzymes: EcoRi and PSTL

4. Buffer: FD Green Buffer

5. 6.0μL of DNA cell stain

6. 5.0μLof plasmid DNA

7. 6.5μL of H2O

  • Chose the following plasmid DNA: BD003, BD004, BD005, BD006
  • We know that the length of BD005 is 2422 bp
  • DPRE also known as D11.12 is 1740bp
  • We assume that BD006 must have the D11.12 construct. If this is so, the length of BD006 would be 4126bp

Results:

  • BD005 and BD006 seemed to be the same length.
  • This could mean that BD006 could be the same plasmid DNA
  • May need to perform new restriction digest.
  • Buffer may have been too diluted. However, this couldn't effect the outcome of size.




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