SBB09 PCR Cloning
From OpenWetWare
Jump to navigationJump to search
Dilute oligonucleotides to be 10 uM
- Add 1 ul of oligo dilution to 9 ul of ddH20
- Mix and spin
Cloning PCR, (products are 937bp & 1600bp, )
- 24 uL water
- 3.3 uL Expand 10x Buffer 2
- 3.3 uL 10x dNTPs (2 mM in each; 0.2 mM final conc)
- 1 uL Oligo Odvs007F (10uM)
- 1 uL Oligo Odvs008rR (10uM)
- 0.5 uL Expand Polymerase 1
- 0.5 uL Template DNA E.coli 0157:H7
- Run this PCR on program 55