PCR: Difference between revisions
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= Colony PCR = | = Colony PCR = | ||
* | * Take colony, mix with 500ul of RO water | ||
* Boiling lysis program | |||
* Then run PCR using [[PCR_Machine| 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 === | === PCR primer design === | ||
Latest revision as of 10:02, 2 May 2015
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/
