IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/07/13

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Entry title

July 13th, 2009

Section 1.

Qubit Fluorometer Assay for Nora's DNA (not PNP-biosensor project)

Notes: DNA concentration in ug/ml. Used protocol as instructed by invitrogen Qubit card. Replaced reagents into Fridge, not Freezer (?)

Tube #

1. name = 346, DNAconc = 212

2. name = 236, DNAconc = 222

3. name = 213, DNAconc = 228

4. name = 186, DNAconc = 173.6

5. name = 49, DNAconc = 222

6. name = 139, DNAconc = 252

7. name = 35, DNAconc = 208

Finished section at 12:45 pm

Section 2: start at 3:30 pm

Primers Dilution

-reconstitute to 100 uM; dilute to 5 uM

1. YGL205WF -> 17.02 nmoles, TE buffer = 100 uM, Amount TE = 170.2 uL

  YGL205WR -> 19.50 nmoles, TE buffer = 100 uM, Amount TE = 195.0 uL
  YJL219WF -> 9.35 nmoles, TE buffer = 100 uM, Amount TE = 93.5 ul

2. YJL219WR -> 19.50 nmoles, TE buffer = 100 uM, Amount TE = 195.0 uL

  YGR287CF -> 12.09 nmoles, TE buffer = 100 uM, Amount TE = 120.9 uL

3. YGR287CR -> 19.76 nmoles, TE buffer = 100 uM, Amount TE = 197.6 uL

  YHL012WF -> 15.40 nmoles, TE buffer = 100 uM, Amount TE = 197.6 uL

4. YHL012WR -> 15.72 nmoles, TE buffer = 100 uM, Amount TE = 157.2 uL

  YDL243CF -> 15.34 nmoles, TE buffer = 100 uM, Amount TE = 153.4 uL

5. YDL243CR -> 16.96 nmoles, TE buffer = 100 uM, Amount TE = 169.6 uL

Note: Saved invitrogen data sheets about these primers. They can be found in the white binder in our lab

Completed dilutions in primer screwtops

PCR

  1. YJL219C
  2. YGR287C
  3. YDL243C
  4. YHL012W
  5. YGL205
  6. -Temp DNA YGL 205 primers
  7. Primers

Template DNA: PFA6alpha GFP/Histag

Primers: See above (07-14 at 3:30)

IGEM 6 conditions : See PCR machine. Start Time: 6:00 p.m. End Time: 10:30 pm

2 beads

0.65 uL template

5 ul of 5 uM primers

37.35 uL H2O



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