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Single Colony PCR to Generate Template for in Vitro Transcription

Single Colony PCR to Generate Template for in Vitro Transcription

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Published by Jackie White
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Categories:Types, Research
Published by: Jackie White on Jun 28, 2013
Copyright:Attribution Non-commercial

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06/28/2013

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Single Colony PCR to Generate Template for In Vitro transcription

For 25 ul reaction: Need per reaction: 6.5 ul GoTaq Master Mix 12.5 ul DEPC H2O 0.25 ul of 20mM pOT for primer 0.25 ul of 20mM pOT rev primer 0.25 ul pfu polymerase (optional) PCR settings: Step 1: 94oC 5 min Step 2: 94oC 45 secs Step 3: 55oC 45 secs + 0.03 sec each cycle (ramping) Step 4: 72oC 1 min 30 sec Repeat steps 2-4 thirty-four times. Hold: 72oC 6 min Run 0.7% agarose gel

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