User:Daniel Goodman/Notebook/Weiss/2010/01/14: Difference between revisions

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Nanodrop on PhyB917 DNA from 7 samples:
!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
|-
|-
|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|-
|}
|}



Revision as of 09:07, 14 January 2010

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Entry title

Will 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:
380|1.88
441|1.87
505|1.87
546|1.85
440|1.88
387|1.85
474|1.86

Taking average, 400 ng/ul

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