Re: [ccp4bb] [OFF-TOPIC] Site-Directed Mutagenesis [OFF-TOPIC]

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Bio Rad "Econo" system
From: narayan viswam nviswam1 {- at -} GMAIL {- dot -} COM
Date: 2012-02-04

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Subject: Re: [OFF-TOPIC] Site-Directed Mutagenesis [OFF-TOPIC]
From: Roger Rowlett rrowlett {- at -} COLGATE {- dot -} EDU
Date: 2012-02-04

Subject: Re: [OFF-TOPIC] Site-Directed Mutagenesis [OFF-TOPIC]
From: Nian Huang huangnian {- at -} GMAIL {- dot -} COM
Date: 2012-02-04

Just a reminder. Quickchange is not PCR. It is linear amplification. It is
very hard to see a band in the gel if you follow the standard protocol.

Nian

On Fri, Feb 3, 2012 at 12:14 PM, Fred wrote:

> Hi CCP4 list,
> Thanks everyone who have answered my post concerning to mutagenesis.
> From quick reading most of the answers, the following seems to be a
> consensus:
> 1) Do not concentrate your PCR product;
> 2) Too much DNA and/or impurities like salts or whatever can inhibits
> transformation;
> 3) Purify your PCR product before transformation if possible or use 3 of 4
> microL of it. This is more or less the amount of DNA showed in the uploaded
> image.
> Kind regards,
> Fred
>
> P.S.: I'll let you know the results.
>

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Bio Rad "Econo" system
From: narayan viswam nviswam1 {- at -} GMAIL {- dot -} COM
Date: 2012-02-04

Next message:
Subject: Re: [OFF-TOPIC] Site-Directed Mutagenesis [OFF-TOPIC]
From: Roger Rowlett rrowlett {- at -} COLGATE {- dot -} EDU
Date: 2012-02-04