1.) Get PCR tubes from last time.
2.) Add 1mL exonuclease I to each tube. Incubate at 37 degrees Celcius for 15 minutes.
3.) Inactivate exonuclease by incubating at 80 degrees Celcius for 15 miutes.
4.) Then label 5 new PCR tubes.
5.) Add 98mL sterile H2O to each tube.
6.) Add 2mL of each sample(PCR from last time plus exonuclease steps) to each new tube. Make sure to use a new pipet tip each time!
7.) Label 5 more PCR tubes.
8.) Add 20mL of Master Mix plus the 2nd primer set to each new tube.
9.) Add 20mL template DNA from diluted PCR sample(from the second set of steps)
4/14/09
*Get PCR samples
*5 little tubes and label (1 per sample)
*Add 5uL loading dye to each tube
*Put 5uL PCR samples into labeled tube; pipet up and down to mix
*Use samples to run on gel (45mins)