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

Divalent Ligand Validation

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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 Empty Size site1 site2 Mass % 6303 EcoRI 1436 EcoRI 1436 100 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.