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elution of dna from gel


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#1 sagar

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Posted 26 April 2009 - 08:43 AM

hi all

how can one improve the extraction quantity of dna from agarose gel manually.

#2 mastermi

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Posted 26 April 2009 - 12:55 PM

Don't use a column.

Give us more information about how you are extracting DNA, and if you want to extract a short or a long DNA fragment...

#3 Felipillo

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Posted 26 April 2009 - 02:16 PM

may be this paper http://dx.doi.org/10...2697(80)90266-3 could help you
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#4 sagar

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Posted 27 April 2009 - 03:43 AM

may be this paper http://dx.doi.org/10...2697(80)90266-3 could help you

Thanks this is of great help.

#5 sagar

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Posted 27 April 2009 - 03:54 AM

Don't use a column.

Give us more information about how you are extracting DNA, and if you want to extract a short or a long DNA fragment...

im carrying out gel elution of a vector of size 2.8kb cut with hindIII and insert fragment of 1.4kb cut with hindIII from low melting agarose gel,i heat it to 65 C and thaw it and use butanol to precipitate it .i also carried out electroelution but the concentration was very less.The starting concentration was 1ug/ul for both vector and insert but now the concentration is less than 50ng/ul

#6 AquaPlasmid

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Posted 14 May 2009 - 10:46 PM

im carrying out gel elution of a vector of size 2.8kb cut with hindIII and insert fragment of 1.4kb cut with hindIII from low melting agarose gel,i heat it to 65 C and thaw it and use butanol to precipitate it .i also carried out electroelution but the concentration was very less.The starting concentration was 1ug/ul for both vector and insert but now the concentration is less than 50ng/ul


Two methods:

1) Use b-agarase digestion (high recovery, needed for library cloning).
2) Melt the gel in a 0.2u spin filter - put in -70 C for 15min - spin (quick but lower yield, but good for most cloning).




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