Team:Calgary/Notebook/Protocols/Process3
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- | <li>After completion of cDNA synthesis, us 1/20th of the first-strand reaction (2-4μL) for PCR amplification in water. Also do 2,3, and 4-fold dilutions of each sample to ensure that the PCRs will be within the primer linear range. Do 3 replicates of each PCR reaction. Run PCRs on a real-time PCR machine (e.g. ABI 7900).</li> | + | <li type="1" value="6">After completion of cDNA synthesis, us 1/20th of the first-strand reaction (2-4μL) for PCR amplification in water. Also do 2,3, and 4-fold dilutions of each sample to ensure that the PCRs will be within the primer linear range. Do 3 replicates of each PCR reaction. Run PCRs on a real-time PCR machine (e.g. ABI 7900).</li> |
Revision as of 20:28, 27 September 2011
Component | Volume |
RNA (40ng – 10ng to 1μg total RNA/rxn) | variable |
Nuclease free water | variable |
qScript cDNA synthesis mix | 4 uL |
Final Volume | 20 uL |