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Team:ETHZ Basel/Biology/Protocols
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(New page: == PCR == 10 ul 5X Buffer 1 ul dNTP's 2 ul Primers (10 fold diluted) 1 ul Template 0.5 ul high fidelity polymerase 35.5 ul water 98°C 30s 98°C 30s 55°C 30s 72°C 60s 52°C 5min 4°C ...)
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(New page: == PCR == 10 ul 5X Buffer 1 ul dNTP's 2 ul Primers (10 fold diluted) 1 ul Template 0.5 ul high fidelity polymerase 35.5 ul water 98°C 30s 98°C 30s 55°C 30s 72°C 60s 52°C 5min 4°C ...)
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Revision as of 13:35, 5 August 2010
Contents |
PCR
10 ul 5X Buffer 1 ul dNTP's 2 ul Primers (10 fold diluted) 1 ul Template 0.5 ul high fidelity polymerase 35.5 ul water
98°C 30s 98°C 30s 55°C 30s 72°C 60s 52°C 5min 4°C
35 cycles
ligation
10 ng storage plasmid 10 times more insert 1 ul ligase buffer (10x T4) fill up water to 10 ul
transformation
chemical:
isolation of plasmids
according to the protocol of NucleoSpin Plasmid (Macherey-Nagel)
agarose gel
1% agarose gel 0.5 g agarose 50 ml 1X TAE heat up with microwave, let cool down a bit 1 ul EtBr pour
