Minnesota/10 July 2008

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| NOTE: Total volume in dephosphorylation = 56.6uL
| NOTE: Total volume in dephosphorylation = 56.6uL
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|2. '''Run RXN model on Calhoun''' (super computer).
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|3. Diluate base vector culture, allow to grow, then miniprep, then streak plates and '''make base vector glycerol stocks.'''
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|4. '''Autoclave dishes'''
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|5.

Revision as of 16:11, 10 July 2008

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1. Dephosphorylation: Continued with double digestion base vector product from 07-09-2008. Used Antartic phosphoylase to dephosphorylate the base vector. Want to dephosphorylate the base vector to prevent it from closing - want it to stay a linear digested product. After dephosphorylation, the base vector was then incubated @ 37C for 30 minutes. Sample was then placed in a heat bath @ 65C for 15 minutes to inactivate enzyme (phosphotase).
Gene 10x Buffer DNA Phosphotase
Base Vector 5.6uL 50.0uL 1.0uL


NOTE: Total volume in dephosphorylation = 56.6uL 2. Run RXN model on Calhoun (super computer). 3. Diluate base vector culture, allow to grow, then miniprep, then streak plates and make base vector glycerol stocks. 4. Autoclave dishes 5.
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