Designing and Preparing dsRNA: Difference between revisions
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==Preparing dsRNA== | ==Preparing dsRNA== | ||
===Template Preparation=== | ===Template Preparation=== | ||
*set up a 250 uL PCR reaction | *set up a 250 uL PCR reaction | ||
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*Dilute RNA 10x in loading dye | *Dilute RNA 10x in loading dye | ||
*Load 1 and 3 uL of diluted RNA, plus 4 or 8 uL of 1 kB Plus DNA Ladder | *Load 1 and 3 uL of diluted RNA, plus 4 or 8 uL of 1 kB Plus DNA Ladder | ||
*Using ImageJ, quantify ladder and dsRNA bands and calculate concentration. Check that the dilutions are close to 3x apart in signal intensity. | *Using ImageJ, quantify ladder and dsRNA bands and calculate concentration. See [[Using ImageJ to Quantify Bands]]. Check that the dilutions are close to 3x apart in signal intensity. | ||
*Label RNA with concentration and store at -20 | *Label RNA with concentration and store at -20 | ||
*see PMID 18388942, PMID 18265350 , PMID 11752672 and http://www.flyrnai.org/DRSC-PRS.html | |||