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Q: Ethanol purification protocol for polymerase chain reaction (PCR) ( No Answer,   2 Comments )
Subject: Ethanol purification protocol for polymerase chain reaction (PCR)
Category: Science > Biology
Asked by: atacdiran-ga
List Price: $15.00
Posted: 28 Jun 2006 14:06 PDT
Expires: 28 Jul 2006 14:06 PDT
Question ID: 741837
What is the protocol for puricifcation of polymerase chain reaction
(PCR) product using ethanol precipitation in order to obtain a PCR
product free from enzymatic contaimants?
There is no answer at this time.

Subject: Re: Ethanol purification protocol for polymerase chain reaction (PCR)
From: mikewa-ga on 30 Jun 2006 13:18 PDT
Is this helpful?
Clean PCR reaction by ethanol precipitation. Make a mix containing
(per sample): 50 ?l 95% Ethanol, 2 ?l 3M NaOAc pH 4.6
Add 52 ?l of the mixture to each well, cover the plate with sealing
mat, and invert 10 times to mix.
Incubate for 15 min at room temperature.
Centrifuge for 30 min at 2000g, remove seal, invert plate onto a stack
of paper towels to drain ethanol.
Add 150 ?l 70% Ethanol.
Centrifuge for 10 min at 2000g. Invert onto paper towels.
Centrifuge inverted plate at 50 g for 1 min.
Let dry plate air dry for 5 min.
Subject: Re: Ethanol purification protocol for polymerase chain reaction (PCR)
From: dops-ga on 27 Jul 2006 11:13 PDT
If the question is for more than just your own edification, you may
want to look into kits that are specifically designed to clean up PCR
rxns. Both Qiagen and Eppendorf make spin kits. They're generally very
easy and much faster than EtoH purifications. Also in my hands, the
resulting product is much clener and there's less loss.

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