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How to isolate specific dna fragment?

Genetics as it applies to evolution, molecular biology, and medical aspects.

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How to isolate specific dna fragment?

Postby ZEGSUS » Mon Aug 22, 2011 6:22 pm

How to isolate specific dna fragment?
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Postby canalon » Mon Aug 22, 2011 7:17 pm

PCR?
Cloning?
Reverse-transcription?
Depends on what you know about your specific fragment, and what you want to do with it. You will need to be more precise if you want a good answer.
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Re: How to isolate specific dna fragment?

Postby ZEGSUS » Mon Aug 22, 2011 7:24 pm

Well I want to isolate specific dna fragment from linear dna which have many dna fragments.I know his code.
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Postby canalon » Mon Aug 22, 2011 7:27 pm

Do a PCR and then separate with gel electrophoresis the fragment of the expected size.
You will have your fragment only, and if you want to clone it, it is easy to add all sorts of modifiers on your primers for further analysis (addition of tags to the protein or the RNA for purification, restriction sites for cloning and expression,...)
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Re: How to isolate specific dna fragment?

Postby ZEGSUS » Wed Aug 24, 2011 8:40 am

And can i use primers for pcr without electrophoresis ?
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Re: How to isolate specific dna fragment?

Postby greatmicrobiologist » Fri Aug 26, 2011 7:33 am

you have to use primers for the specific sequence required in PCR. Then for the separation of your required fragment form the bulk of fragments you have to go for electrophoresis, i.e. purification.
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Re: How to isolate specific dna fragment?

Postby JackBean » Fri Aug 26, 2011 12:18 pm

ZEGSUS wrote:And can i use primers for pcr without electrophoresis ?


You must use primers for PCR, no matter, what you do with the reaction afterwards ;) You can e.g. purify the amplicon by some spin columns or directly digest with RE and afterwards purify etc. ;)
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Re: How to isolate specific dna fragment?

Postby merv » Mon Oct 03, 2011 2:16 am

greatmicrobiologist wrote:you have to use primers for the specific sequence required in PCR. Then for the separation of your required fragment form


think you meant from not form

greatmicrobiologist wrote: the bulk of fragments you have to go for electrophoresis, i.e. purification.


i think we both know that separation by electrophoresis is not the same as purification. :roll:
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