User:DrJones1935/16 July 2012
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(Created page with "====I. Ordered Cross-over PCR Primers from Keck==== *Primers were ordered with the name/sequence in the following table: {| class="wikitable" |- ! Name !! Tm (Tm overhang/Tm ann...")
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(Created page with "====I. Ordered Cross-over PCR Primers from Keck==== *Primers were ordered with the name/sequence in the following table: {| class="wikitable" |- ! Name !! Tm (Tm overhang/Tm ann...")
Newer edit →
Revision as of 23:21, 3 October 2012
I. Ordered Cross-over PCR Primers from Keck
- Primers were ordered with the name/sequence in the following table:
Name | Tm (Tm overhang/Tm anneal) | Length (bp) | Sequence (overCUTSITEanneal) |
---|---|---|---|
cT5_pAf | 66.64 (25.23/58.79) | 30 (10/20) | 5'-tgacTCTAGAtttgctttgtgagcggataa-3' (XbaI) |
cT5_pAr | 69.08 (30.28/55.61) | 25 (10/15) | 5'-gtaaCTGCAGgagagcgttcaccga-3' (PstI) |
- See Project Page for more info
II. Resuspend Lyophilized Primers for Cross-over PCR
- Spin down tubes
- Add the volume specified in the table of TE Buffer to the primer tubes
Name | Volume (uL) |
---|---|
p1f | 208.35 |
p1r | 217.78 |
p2f | 199.75 |
p2r | 199.6 |
p3f | 199.75 |
p3r | 190.55 |
p4f | 248.1 |
p4r | 149.85 |
p5f | 127.75 |
p5r | 199.65 |
- Vortex all tubes briefly
- Spin all tubes down
- Incubate on bench for 10 minutes
- Store at -20 oC