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

iGEM Project name 1

Entry title

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

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