The two MspI restriction digestions from earlier today for our project with Qiagen were subjected to phenol:chloroform cleanup and subsequent ethanol precipitations.
Phenol:chloroform clean up procedure:
- Added equal volume (50μL) of phenol:chloroform:IAA (25:24:1) to each sample.
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Vortexed.
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Centrifuged 5mins, 16,000g at room temperature.
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Transferred aqueous phase (top layer) to clean 0.5mL snap-cap PCR tube.
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Added equal volume of chloroform (50μL) to aqueous phase.
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Vortexed.
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Centrifuged 5mins, 16,000g at room temperature.
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Transferred aqueous phase (top layer) to clean 0.5mL snap-cap PCR tube.
Performed ethanol precipitation on both samples according to lab protocol.
Resuspended precipitated DNA in 25μL Buffer EB (Qiagen).
Will quantify with Qubit 3.0.
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