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Team:TU Darmstadt/protocols/PCR
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<br> | <br> | ||
<B> Chemicals & consumables <br></B> | <B> Chemicals & consumables <br></B> | ||
| - | DNA | + | DNA template<br> |
Nuclease-free water<br> | Nuclease-free water<br> | ||
Primer<br> | Primer<br> | ||
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<br> | <br> | ||
<B> Procedure<br></B> | <B> Procedure<br></B> | ||
| - | + | This is a commonly used protocol thta we used for Taq-polymerase<br> | |
| - | PCR program (30 cycles) | + | <B>PCR program (30 cycles)<br></B> |
initial denaturation 95°C, 60s | initial denaturation 95°C, 60s | ||
denaturation 95°C, 45s | denaturation 95°C, 45s | ||
| - | annealing 54,5°C, 90s | + | annealing 54,5°C, 90s |
elongation 72°C, 120s | elongation 72°C, 120s | ||
final elongation 72°C, 120s | final elongation 72°C, 120s | ||
| - | run | + | After worth run an analytic 1% agarose gel and purificate the product with Wizard SV Gel and PCR Clean-Up System (Promega) |
<br> | <br> | ||
Revision as of 16:43, 4 October 2013
PCR
Short report
Materials
Equipment
Micropipettes with sterile tips
PCR machine
PCR tubes
Chemicals & consumables
DNA template
Nuclease-free water
Primer
Polymerase buffer
dNTP mix
DMSO
Procedure
This is a commonly used protocol thta we used for Taq-polymerase
PCR program (30 cycles)
initial denaturation 95°C, 60s
denaturation 95°C, 45s
annealing 54,5°C, 90s
elongation 72°C, 120s
final elongation 72°C, 120s
After worth run an analytic 1% agarose gel and purificate the product with Wizard SV Gel and PCR Clean-Up System (Promega)
