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
 * 2) Add 1µl BamHI
 * 3) Add 1µl XhoI
 * 4) 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


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