PCR

Colony PCR

• Take colony, mix with 500ul of RO water
• Boiling lysis program
• Then run PCR using Techne TC-312
• M13 primers (LacZ region of PUC19) and S16 primers (universal bacteria)
• COL program. 30 cycles. Annealing is 53C (ideal for S16, 56 would be better for M16). Hold at 4

PCR primer design

• http://www.cybertory.org/exercises/primerDesign/index.html
• http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html
• http://www.premierbiosoft.com/primerdesign/index.htm
• http://molbiol-tools.ca/PCR.htm
• http://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?LINK_LOC=BlastHome
• http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi
• http://bibiserv.techfak.uni-bielefeld.de/genefisher2/