"
Team:University College London/Protocols/PCR
From 2012.igem.org
(Difference between revisions)
Rwilkinson (Talk | contribs) (Created page with "<noinclude>{{:Team:University_College_London/templates/head|coverpicture=week4}} == Transformation Protocol 2 == </noinclude> '''Step 1 - Setting up PCR tubes:''' Thaw reagents...") |
m (moved Team:University College London/LabBook/Protocols/PCR to Team:University College London/Protocols/PCR: consistency) |
||
| (2 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
<noinclude>{{:Team:University_College_London/templates/head|coverpicture=week4}} | <noinclude>{{:Team:University_College_London/templates/head|coverpicture=week4}} | ||
| - | == | + | == PCR == |
</noinclude> | </noinclude> | ||
| Line 45: | Line 45: | ||
| - | |||
<noinclude> | <noinclude> | ||
{{:Team:University_College_London/templates/foot}} | {{:Team:University_College_London/templates/foot}} | ||
</noinclude> | </noinclude> | ||
Latest revision as of 15:41, 22 August 2012
PCR
Step 1 - Setting up PCR tubes: Thaw reagents and add to PCR tubes in the proportions described in the table below
| PCR Components | Volume (ul) |
|---|---|
| 5x Reaction Buffer | 10 |
| 25mM MgCl2 | 4 |
| 10mM dNTPs | 1 |
| 10uM Forward primer | 5 |
| 10uM Reverse primer | 5 |
| DNA Polymerase | 0.25 |
| Nuclease Free Water | 24.25 |
| Template DNA | 0.5 |
| Total Volume | 0.5 |
Step 2 - PCR program: Add PCR tubes to a thermocycler and run under the following conditions.
| PCR conditions | Temp (oC) | Time (s) |
|---|---|---|
| Initial Denaturation (1 cycle) | 95 | 30 |
| Denaturation/Annealing/Extension (30 cycles) | 95/55/72 | 10/25/120 |
| Final Extension (1 cycle) | 72 | 600 |
| Hold | 4 | ∞ |
