5 replies to this topic

[sagar]

hi all

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

[mastermi]

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...

[Felipillo]

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

[sagar]

Felipillo, on Apr 27 2009, 03:46 AM, said:

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

Thanks this is of great help.

[sagar]

mastermi, on Apr 27 2009, 02:25 AM, said:

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

[AquaPlasmid]

sagar, on Apr 27 2009, 04:54 AM, said:

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).