Preparing samples for sequencing at UNC

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DNA Sample Preparation for Sequencing at UNC
Miniprep DNA
1. RNAse – either use TE with RNAse to resuspend pellet or add 0.5 l RNAse.
2. Prepare a 0.5 ml tube with 0.7 g DNA + 10 pmol primer (0.2 l of our standard
concentration) in 20 l total volume. Usually, 2-5 l of miniprep DNA per reaction is
sufficient.
PCR product
1. Cut single band out of gel and spin through glass wool.
2. Clean DNA with Invitrogen PureLink PCR cleanup kit.
3. Prepare a 0.5 ml tube with 10 ng/100 bp DNA + 10 pmol primer (0.2 l of our standard
concentration) in 20 l total volume.
Sequence request form
1. The sequence request form can be accessed from the yellow “Sequence” bar.
Fill out the form; print one copy; save a copy under another file name.
Updated 2/27/08 by Matt LaFave
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