User:Robwarden/Notebook/Divalent Ligand Validation/2009/02/27

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pMSCVN-H1C Validation

Expected Banding Patterns

Correct
Size site1 site2 Mass %
6587 EcoRI 5834 EcoRI 1720 62
1838 EcoRI 1720 EcoRI 3558 17
1508 EcoRI 4326 EcoRI 5834 14
768 EcoRI 3558 EcoRI 4326 7
Image:PMSCVN-H1C-EcoRI.png
Empty
Size site1 site2 Mass %
6303 EcoRI 1436 EcoRI 1436 100
Image:PMSCVN-EcoRI.png

Miniprep

EcoRI Digest

 10 μL DNA
  2 μL 10X Buffer EcoRI ->   26 μL
 .5 μL EcoRI            ->  6.5 μL
7.5 μL MilliQ           -> 97.5 μL

Incubate at 37°C for 1hr.

Results

  • Clearly empty vectors for most clones.
  • α2,3,6 had a strong band at ~2.2kb (maybe the 1.5kb & .7kb bands?). Faint bands at 6.5kb and 1.8kb. Liquid cultures restarted by Lu. Will be sent for sequencing on Monday 4/2/09.
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