179
edits
DNA extraction and precipitation with phenol-chloroform: Difference between revisions
From London Hackspace Wiki
DNA extraction and precipitation with phenol-chloroform (view source)
Revision as of 20:11, 2 October 2016
, 2 October 2016Grammar
(Created page with "== Phenol-chloroform extraction == The process is dividing into three phases: homogenisation, extraction, and resuspension. === Homogenisation === The first step is to brea...") |
(Grammar) |
||
| Line 1: | Line 1: | ||
== Phenol-chloroform extraction == | --[[User:Wzdd|Wzdd]] ([[User talk:Wzdd|talk]]) 21:11, 2 October 2016 (BST)== Phenol-chloroform extraction == | ||
The process is | The process is divided into three phases: homogenisation, extraction, and resuspension. | ||
=== Homogenisation === | === Homogenisation === | ||
The first step is to break open the cells and suspend their contents in an extractable form. | The first step is to break open the cells and suspend their contents in an extractable form. | ||
# If | # If your sample is cells in liquid (e.g. blood, spit), pelletise the sample by centrifuging. Discard the liquid. | ||
# Suspend in 100ul homogenisation buffer, comprising 10mm edta, 60mm nacl, and 10mm tris (lowered to a pH of 7.5 using HCl) | # Suspend in 100ul homogenisation buffer, comprising 10mm edta, 60mm nacl, and 10mm tris (lowered to a pH of 7.5 using HCl) | ||
# Add equal volume solubilisation buffer, comprising 2% SDS, 30mm EDTA, 200mm tris (lowered to a pH of 9 using HCl) | # Add equal volume solubilisation buffer, comprising 2% SDS, 30mm EDTA, 200mm tris (lowered to a pH of 9 using HCl) | ||
