Team:Northwestern/pcr
From 2013.igem.org
(Difference between revisions)
2013.igem.nu (Talk | contribs)
(Created page with "{{:Team:Northwestern/Templates/Skinning}} <html> <style> p { color:black; font-family: helvetica; } .container { display: inline-block; background-color: #C3...")
Newer edit →
(Created page with "{{:Team:Northwestern/Templates/Skinning}} <html> <style> p { color:black; font-family: helvetica; } .container { display: inline-block; background-color: #C3...")
Newer edit →
Revision as of 23:57, 27 September 2013
PCR Amplification
To a small PCR tube (on ice) add:
- enough water to fill to 50 ul (19 uL)
- 1 ul of template DNA
- 2.5 ul each of 10 uM forward and reverse primers
- 25 ul of PCR Master Mix
Quickly transfer PCR tubes from ice to a preheated thermocycler with the following conditions:
- initial denaturation: 94°C for 30 seconds
- 25-35 cycles: 94°C for 15-30 seconds (we did 30 sec)
- 45-68°C for 15-60 seconds (we did 60°C for 30 sec)
- 68°C for 1min/kb (we did 2 min)
- final extension: 68°C for 5 min
- hold: 4°C for as long as necessary
NOTE: For colony PCR just add a single colony using a pipette tip as the last thing you add to the solution instead of the template DNA. Twirl the colony into the solution in the tube and then proceed as usual