Team:Chiba/protocol/PCR

From 2008.igem.org

(Difference between revisions)
(New page: >Protcol PCR)
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PCR
PCR
 +
 +
*Resuspend primer in Nuculease free water to 100 µM
 +
 +
*PCR mix
 +
#DNA template          1µL
 +
#Fwd primer            10µL (final con. 10 pM)
 +
#Rev primer            10µL
 +
#10x thermo pol buffer 10µL
 +
#dNTP mix              10µL
 +
#DNA pol.              1µL
 +
#dH20                  58µL
 +
<br>----------------------------
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<br>                  100µL
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*PCR cycle
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* Start: 94 °C for 5 min. (melt)
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* cycle:          melt:            1 min.
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*              anneal :          30 sec.
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* cycle end: extension:72 °C for 3.5 min.
 +
* 25 cycles
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* 72 °C for 10 min
 +
* End: keep at 6 °C forever

Revision as of 02:10, 11 September 2008

>Protcol

PCR

  • Resuspend primer in Nuculease free water to 100 µM
  • PCR mix
  1. DNA template 1µL
  2. Fwd primer 10µL (final con. 10 pM)
  3. Rev primer 10µL
  4. 10x thermo pol buffer 10µL
  5. dNTP mix 10µL
  6. DNA pol. 1µL
  7. dH20 58µL


----------------------------
100µL

  • PCR cycle
  • Start: 94 °C for 5 min. (melt)
  • cycle: melt: 1 min.
  • anneal : 30 sec.
  • cycle end: extension:72 °C for 3.5 min.
  • 25 cycles
  • 72 °C for 10 min
  • End: keep at 6 °C forever
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