elution of dna from gel - (Apr/26/2009 )
hi all
how can one improve the extraction quantity of dna from agarose gel manually.
-sagar-
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...
-mastermi-
may be this paper http://dx.doi.org/10.1016/0003-2697(80)90266-3 could help you
-Felipillo-
Felipillo on Apr 27 2009, 03:46 AM said:
may be this paper http://dx.doi.org/10.1016/0003-2697(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...
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
-sagar-
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).
-AquaPlasmid-