User:Howard Boland/Notebook/Art from Synthetic Biology/2010/11/19

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Overnight Growth

All 4 tubes (10ml) grew.

Miniprep

I ran a miniprep and eluted each sample in 50µl of H2O.

Digestion pSense66 PstI and XhoI/BamHI

2xMaster Mix

  1. 1µl 100xBSA
  2. 10µl 10xNEB#3
  3. 25µl H2O
  4. 60µl pSense66

For each of two reaction tubes add 48µl Master Mix

  1. Tube 1
    1. Add 1µl BamHI
    2. Add 1µl XhoI
  2. Tube 2: 2µl PstI

Incubate in 37ºC for 2 hours

Gel Check

I set up a gel to check the miniprep and the digestion results.

  1. Lane 1: 10µl 1kb NEB Quick Ladder
  2. Lane 2: 50µl pSense66 PstI Digestion (genomic contamination or other error)
  3. Lane 3: BLANK
  4. Lane 4: 50µl pSense66 XhoI/BamHI Digestion (genomic contamination or other error)
  5. Lane 5: 5µl Miniprep purification of overnight growth
  6. Lane 6: 5µl Miniprep purification of overnight growth
  7. Lane 7: 5µl Miniprep purification of overnight growth
  8. Lane 8: 5µl Miniprep purification of overnight growth

Gel Picture 19112010-plasmids-and-cuts.jpg