Difference between revisions of "PCR"

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= Colony PCR =
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* Take colony, mix with 500ul of RO water
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* Boiling lysis program
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* Then run PCR using [[PCR_Machine| Techne TC-312]]
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* M13 primers (LacZ region of PUC19) and S16 primers (universal bacteria)
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* COL program. 30 cycles. Annealing is 53C (ideal for S16, 56 would be better for M16). Hold at 4
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=== PCR primer design ===
 
=== PCR primer design ===
 
* http://www.cybertory.org/exercises/primerDesign/index.html
 
* http://www.cybertory.org/exercises/primerDesign/index.html

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

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