User:Daniel Goodman/Notebook/Weiss/2010/01/14: Difference between revisions
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Water 12.82857143 | Water 12.82857143 | ||
AVRII | ||AVRII | ||
Volume of DNA|0.571428571 | Volume of DNA|0.571428571 | ||
Volume of enzyme|0.1 | Volume of enzyme|0.1 | ||
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Water|12.82857143 | Water|12.82857143 | ||
BSA|0 | BSA|0 | ||
Total Volume|15 | Total Volume|15|| | ||
Revision as of 09:01, 14 January 2010
 Project name
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Entry titleWill Run PhyB917, PhyB, and PIF3 on a Gel PhyB917 Cut with BstBI - two bands, 3579 and 5314 incubate at 65 degrees with NEBuffer 4 Nanodrop on PhyB917 DNA from 7 samples: 1 380,1.88 2 441, 1.87 3 505, 1.87 4 546 1.85 5 440 1.88 6 387 1.85 7 474 1.86
Calculate digestion mix for 7.25 aliquots: One enzyme digest calculation based on mass of DNA to be digested Mass of DNA / ug 0.2 Units of enzyme 0.2 Concentration of DNA ng/ul 400 Concentration of enzyme Units/ul 20 Overdigestion factor 2 Volume of DNA / ul 0.5 Volume of enzyme / ul 0.02 Calculate Total Volume 0.5 Actual Total Volume 15 NEB Buffer 1.5 Water 12.98 BSA 0 Water 12.98 ENZYME 0.145 BUFFER(4) 10.875 WATER 94.105 BSA 0 (none req'd for BstBII) TOAL VOL PER ALIQ: 14.5 (add 0.5 of dna to each) PIF3 (possibly) Cut with AvrII - two bands, 2164 and 4791 incubate at 37 deg w/ N EBuffer 4 Nanodrop on PIF3 DNA sample: 357.4 ng/ul One enzyme digest calculation based on mass of DNA to be digested Mass of DNA / ug 0.2 Units of enzyme 0.2 Concentration of DNA ng/ul 350 Concentration of enzyme Units/ul 4 Overdigestion factor 2 Volume of DNA / ul 0.571428571 Volume of enzyme / ul 0.1 Calculate Total Volume 1 Actual Total Volume 15 NEB Buffer 1.5 Water 12.82857143 BSA 0 Water 12.82857143 |
AVRII
Volume of DNA|0.571428571 Volume of enzyme|0.1 NEB Buffer|1.5 Water|12.82857143 BSA|0 Total Volume|15||
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