Protocols for DNA extraction

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Protocols for DNA extraction

• Park, Woosin
• Beel, K-JIST

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• Sample requirements
• 1. The sample is need as much as at least 0.03g
  in wet weight

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• Reagents
• 1. Solution I SDS ? breakdown of cell
  membrane, ?? ???? ??????? ???? ??? ?? ?? ????.
• 2. Solution II benzene chloride
• 3. Solution III NaAc
• All the reagents are made in WACO
• company

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• Procedure

Injecting 300 µl of solution I
Removing the liquid phase
Concentrating sample by centrifugation, 12kg and
4oC for 10 min
Injecting 150 µl of solution II
Vortex the sample
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Injecting 150 µl of solution III
Ice
Incubation for 15min at 50 oC
Leave the sample on ice for 15 min
When moving the tube from centrifuge, be
cautious for the band not to be suspended.
DNA is included in the supernatant
Centrifuge the sample in 12kg and 4 oC for 15
min
Protein band
Move the supernatant into a new tube (1.5 ml)
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Move the supernatant into a new tube (1.5 ml)
Centrifuge the sample in order to remove unknown
contaminants
Add 100 ethanol 2-2.5 times as the liquid sample
Remove the supernatant
Centrifuge the sample immediately in 12kg and 4
oC for 10 min
Mix the sample gently
DNA
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Add about 0.2 ml of 70 ethanol in order to wash
the DNA sample
Remove the supernatant
Centrifuge the sample immediately in 12kg and 4
oC for 10 min
Add 0.05ml of TE (Tris-EDTA, pH8.0), split the
sample into 5 tubes in 10 µl respectively and
store all the samples at -20 oC
Take off the cap and dry in the air for 30 min