Team:Calgary/Notebook/Protocols/Process3
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<li type="1">Thaw all components (except the enzyme/buffer mix), mix thoroughly and centrifuge before use.</li> | <li type="1">Thaw all components (except the enzyme/buffer mix), mix thoroughly and centrifuge before use.</li> | ||
- | <li>Add the following to a 0.2mL thin-walled PCR tube sitting on ice:</li> | + | <li type="1">Add the following to a 0.2mL thin-walled PCR tube sitting on ice:</li> |
TITLE=cDNA Synthesis/Reverse Transcription Reaction| | TITLE=cDNA Synthesis/Reverse Transcription Reaction| | ||
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<div style="margin-bottom:60px"> | <div style="margin-bottom:60px"> | ||
<table width="700"> | <table width="700"> | ||
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</table> | </table> | ||
- | <li>Thaw all components (except enzyme), mix thoroughly and centrifuge before use.</li> | + | <li type="1">Thaw all components (except enzyme), mix thoroughly and centrifuge before use.</li> |
- | <li>Mix components by gently vortexing and then centrifuge 4s to collect contents.</li> | + | <li type="1">Mix components by gently vortexing and then centrifuge 4s to collect contents.</li> |
- | <li>Incubate in a PCR machine using the following conditions:</li> | + | <li type="1">Incubate in a PCR machine using the following conditions:</li> |
<div style="margin-bottom:60px"> | <div style="margin-bottom:60px"> |
Revision as of 20:24, 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 |
5 min at 25°C | |
30 min at 42°C | |
5 min at 85°C | variable |
Hold at 4°C |