118
edits
DNA extraction - Salting Out: Difference between revisions
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→Procedure
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==Procedure== | ==Procedure== | ||
Tissue Digestion | *Tissue Digestion | ||
#Add 5μL Proteinase K to each mL of Digestion Buffer (final 0.5mg/mL) | #Add 5μL Proteinase K to each mL of Digestion Buffer (final 0.5mg/mL) | ||
#Homogenise (or simply place) tissue in solution | #Homogenise (or simply place) tissue in solution | ||
| Line 31: | Line 31: | ||
#Transfer supernatant into a new tube | #Transfer supernatant into a new tube | ||
Precipitation of Protein and Cell Debris | *Precipitation of Protein and Cell Debris | ||
#Add 1/10 volume of Sodium Acetate 3M pH 5.2 (final 0.3M) | #Add 1/10 volume of Sodium Acetate 3M pH 5.2 (final 0.3M) | ||
#Invert to mix and incubate at -20°C for ~15 minutes | #Invert to mix and incubate at -20°C for ~15 minutes | ||
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#Transfer supernatant to a new tube | #Transfer supernatant to a new tube | ||
Precipitation of DNA | *Precipitation of DNA | ||
#Add >2 volumes of 98% ethanol (final 60-80%) | #Add >2 volumes of 98% ethanol (final 60-80%) | ||
#Invert to mix and incubate at -20°C for ~15 minutes | #Invert to mix and incubate at -20°C for ~15 minutes | ||
