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--- archive:lab_protocols:solid_run1 [2010/03/30 04:44]
hyjkim created
+++ archive:lab_protocols:solid_run1 [2015/09/02 16:57] (current)
ceisenhart ↷ Page moved from lab_protocols:solid_run1 to archive:lab_protocols:solid_run1
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 ===== Solid Sequencing Run 1 =====
 ==== Meta ====
+Prepared by: Eveline Hesson
 Date:
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 ==== DNA Extraction ====
 ==== Library Preparation ====
+This library was prepared by following ABI's standard 2x25 mate-pair library preparation protocol. {{:lab_protocols:solid3_libprep_guide.pdf|}}
+Detailed notes of the protocol can be found in the following pdf: {{:lab_protocols:solid-mate-pair-notes.pdf|}}
+=== Shearing DNA ===
+Extracted genomic DNA was sheared using a [[https://products.appliedbiosystems.com/ab/en/US/adirect/ab?cmd=catNavigate2&catID=604432|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 ====

Banana Slug Genomics