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archive:lab_protocols:solid_run1

Solid Sequencing Run 1

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Prepared by: Eveline Hesson

Date:

Overview:

DNA Extraction

Library Preparation

This library was prepared by following ABI's standard 2×25 mate-pair library preparation protocol. solid3_libprep_guide.pdf

Detailed notes of the protocol can be found in the following pdf: solid-mate-pair-notes.pdf

Shearing DNA

Extracted genomic DNA was sheared using a Hydroshear to an approximate size of 4-5kb. The shearing was done using Standard_Assembly, sc 15, 5 cycles for 150 µl total volume.

Total recovered DNA after hydroshear was 217 µl or 26 µg.

Repair the DNA Ends

The DNA ends of sheared gDNA were blunt ended and phosphorylated using Epicentre's End-It DNA repair Kit. 217 µl of sheared gDNA was reacted in a total volume of 400 µl according to manufacturer's standards. The reaction was incubated for 30 minutes at room temperature.

Purify the End-Repaired DNA using the QIAquick Gel Extraction Kit

Emulsion PCR

Sequencing Data

Discussion

, 2010/04/06 16:08

Who did each of the steps of the protocol? dates?

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archive/lab_protocols/solid_run1.txt · Last modified: 2015/09/02 09:57 by ceisenhart