Team:Bielefeld-Germany/Labjournal/Molecular
From 2013.igem.org
PCR Protocols and Programs
Phusion 50 µl
Reagent | 1x | 8x |
---|---|---|
Hf-Buffer 5x | 10.0 | 88.0 |
dNTPs (10mM) | 2.0 | 17.6 |
Primer1 (10mM) | 1.0 | 8,8 |
Primer2 (10mM) | 1.0 | 8,8 |
Template (16.5ng) | 3.0 | 26.4 |
DMSO | 4.0 | 35.2 |
Phusion Polymerase (0.5u) | 1.0 | 8.8 |
H2O | 26.0 | 228.8 |
Program Phusion | ||||
---|---|---|---|---|
98 °C | 98 °C | 68 °C | 72 °C | 72 °C |
30s | 10 s | 30 s | 1:35 | 6:00 |
35 cycles of steps 2 to 4 |
Taq 25 µl
Reagenz | 1x | 8x |
---|---|---|
Buffer 5x | 5,0 | 44 |
MgSO4 | 0 | 0 |
dNTPs | 2,5 | 22 |
Primer1 | 1,0 | 8,8 |
Primer2 | 1,0 | 8,8 |
Template | 1,5 | 13,2 |
DMSO | 2,0 | 17,6 |
Polymerase | 0,5 | 4,4 |
H2O | 11,5 | 101,2 |
Program Taq | ||||
---|---|---|---|---|
98 °C | 98 °C | x °C | 72 °C | 72 °C |
2 min | 30 s | 30 s | y | 6 min |
x: according to Primer Tm y: 15s/kb |