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DNA extraction from tissue: Difference between revisions
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Protocol for extraction of DNA from tissue or embryos. | Protocol for extraction of DNA from tissue or embryos. | ||
== | == Proteinase K digestion == | ||
#Mix DNA extraction buffer | #Mix DNA extraction buffer | ||
#*98 μl [[Oneill_Lab:Chemicals#ReagentB|ReagentB]] | #*98 μl [[Oneill_Lab:Chemicals#ReagentB|ReagentB]] | ||
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#Incubate at 50°C overnight | #Incubate at 50°C overnight | ||
==PCI== | == Phenol/chloroform/isoamyl (PCI) == | ||
#Prepare PCI mix | #Prepare PCI mix | ||
##One part tris-saturated phenol to one part 24:1 Chloroform:Isoamyl alcohol | ##One part tris-saturated phenol to one part 24:1 Chloroform:Isoamyl alcohol | ||
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#Continue to precipitation | #Continue to precipitation | ||
==Ethanol | == Ethanol precipitation == | ||
#Add 2 volumes 100% EtOH | #Add 2 volumes 100% EtOH | ||
#Add 1/10 volume 3M Sodium Acetate pH 5.0 | #Add 1/10 volume 3M Sodium Acetate pH 5.0 | ||
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* [[Image:2stars.png]] [http://www.bio.com/protocolstools/protocol.jhtml?id=p1689 DNA extraction from tail or tissue] - at BioProtocol from unknown contributor | * [[Image:2stars.png]] [http://www.bio.com/protocolstools/protocol.jhtml?id=p1689 DNA extraction from tail or tissue] - at BioProtocol from unknown contributor | ||
* [http://www.protocol-online.org/prot/Molecular_Biology/DNA/DNA_Extraction___Purification/DNA_Extraction_from_Cell_and_Tissue/ Protocol Online links to DNA extraction protocols] | * [http://www.protocol-online.org/prot/Molecular_Biology/DNA/DNA_Extraction___Purification/DNA_Extraction_from_Cell_and_Tissue/ Protocol Online links to DNA extraction protocols] | ||
