User:Howard Boland/Notebook/Art from Synthetic Biology/2010/09/10

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Overnight Growth Result

All 6 samples grew well. To ensure I had sufficient original stock, I made 3 tubes of glycerol stock of pUA66 and stored at -80ºC.

Streaking Result

Both plates grew. However, none of the colonies on the pUA66katE#2, Blue exhibited visible expression of GFP.

Glycerol stock:

  1. 700µl pUA66
  2. 300µl 50% Glycerol

The remaining 5 samples were prepared for miniprep as I already had stock of these

Miniprep

Digestion

I digested 3 samples of different stock using 15µl of DNA.

Mastermix:

  1. ---to come

Gel Result

I ran both the 5 miniprep samples (2µl each) and the 3 digested samples on two different gels.

The Miniprep samples were run on a 1% gel to check for genomic contamination - none where found but the position of the bands surprised me.

  1. 5µl 1kb NEB Quick Ladder
  2. 2µl pUA66katE#2, Blue
  3. 2µl pUA66katE#2, Blue
  4. 2µl pUA66katE#2, Green
  5. 2µl pUA66katE#2, Green
  6. 2µl pUA66katE#2, Black

The 3 digested samples where run on a 2% gel with a 100bp ladder.

  1. 5µl 100bp NEB Ladder
  2. 30µl pUA66katE#2, Blue
  3. 30µl pUA66katE#2, Green
  4. 30µl pUA66katE#2, Black

Fluorescent Microscopy outcomes

Some background flourescent was observed


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