Friday, December 16, 2005
DNA/RNA Blotting With the Trans-Blot® SD Semi-Dry Electrophoretic Transfer Cell
The Trans-Blot SD DNA/RNA agarose gel support frame, used with the Trans-Blot
SD semi-dry electrophoretic transfer cell, allows blotting of DNA/RNA
from agarose gels to nylon membrane in minutes, without any gel pretreatments.
The Trans- Blot SD semi-dry electrophoretic transfer cell generates a
high field strength (V/cm2) which facilitates rapid transfers. The blotting
frame allows highly efficient transfers by eliminating crushing of agarose
gels.
DNA Transfers in 10 Minutes
PCR* fragments, plasmid, and vector DNA ranging in size from several hundred
bases to 15 kilobases can now be quickly transferred in as little as 10
minutes. Set-up time is reduced to just minutes because hydrolyzation
of DNA fragments is not required prior to transfer.
RNA Transfers in 30–35 Minutes
The agarose gel support frame can also be used to transfer RNA up to 3.5
kb in 30–35 minutes, although transfer of 28s rRNA is equally efficient
as determined by EtBr staining.1
Northern and Southern Blotting
The agarose gel support frame will accommodate 6 mm thick agarose gels
up to 15 x 20 cm. Table 1 shows the volumes of agarose required for each
gel size.
For convience, 10x TBE and TAE premixed buffers are avaliable. Extra
thick, 2.6 mm, filter paper is necessary to provide enough buffer for
transfer. The extra thick blot paper must be identical to the dimensions
of the agarose gel. Bio-Rad’s extra thick blot paper is available in four
sizes: 7.5 x 10 cm, 10 x 15 cm, 15 x 15 cm, and 15 x 20 cm. Use of any
other type of blot paper will decrease transfer efficiency.
References
1. Lane, K., Zoia, O. and Davidson, J. M., Biotechniques, 12, 3,
340–346 (1992).
2. Trans-Blot Instruction Manual.
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