User:Howard Boland/Notebook/Art from Synthetic Biology/2010/11/23

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Check plate overnight growth with H2O2 disk check
The plate showed growth only around 1/1000 and 1/10000 dilution of 33% H2O2 concentration but I was not observing GFP under UV-light. The following concentrations can be observed by reading the disk starting with the one below the arrow and reading clockwise.
 * 1) Disk 1: 1/1
 * 2) Disk 2: 1/10
 * 3) Disk 3: 1/100
 * 4) Disk 4: 1/1000
 * 5) Disk 5: 1/10000
 * 6) Centre Disk: Water / Control



Digestion pUA66 and katE with PstI
2xMaster Mix
 * 1) 1µl 100xBSA
 * 2) 10µl 10xNEB#3
 * 3) 4µl PstI
 * 4) 25µl H2O

For each of two reaction tubes add 20µl Master Mix
 * 1) Tube 1: 30µl pUA66
 * 2) Tube 2: 20µl katE

Incubate in 37ºC for 2 hours

Digestion pSense66 XhoI/BamHI

 * 1) 0.5µl 100xBSA
 * 2) 5µl 10xNEB#3
 * 3) 1µl XhoI
 * 4) 1µl BamHI
 * 5) 12.5µl H2O
 * 6) 30µl pSense66

Incubate in 37ºC for 2 hours

Gel of Digestion

 * 1) Lane 1: 10µl 1kb Quick Ladder
 * 2) Lane 2: 10µl pUA66katE - PstI
 * 3) Lane 3: 50µl pUA66 - PstI
 * 4) Lane 4: BLANK
 * 5) Lane 5: 50µl katE - PstI
 * 6) Lane 6: BLANK
 * 7) Lane 7: 50µl pSense66 - XhoI / BamHI
 * 8) Lane 8: Blank

Gel Picture

Digestion pSense66 XhoI/BamHI

 * 1) 0.5µl 100xBSA
 * 2) 5µl 10xNEB#3
 * 3) 1µl XhoI
 * 4) 1µl BamHI
 * 5) 12.5µl H2O
 * 6) 30µl pSense66

Incubate in 37ºC overnight, for 16 hours


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