|
Transformation of E.Coli is still essential in retrieving the recombinant Plasmid DNA clone of choice.
Since the introduction (Cohen 1972) of the CaCl2 method the transformation efficacy increased
enormously and was improved in 1983 by Hanahan. With the introduction of prepacked competent cells
another significant improvement was made.
We at Synvolux developed a TRAnsformation ENhancer for E. coli strains.
The use of TRAEN will increase the amount of colonies retrieved. Depending on the plasmid length
( 3,5 ~ 10 Kb ) and purification state of the plasmid the transformation efficacy will increase by
200% and more. In a typical experiment with a 5 Kb plasmid we retrieved 5 times more colonies
using TRAEN in the transformation of commercially available competent cells. Instead of using prepacked
competent cells with TRAEN, the same results were obtained when we used cells prepared with the
CaCl2 method in combination with TRAEN.
# cfu per 10 ng plasmid DNA (cfu= colony forming units)
Standard transformation 100
0,3 ul TRAEN transformation 356
0,5 ul TRAEN transformation 350
In ligation experiments 10 ng DNA ligation mix was plated, the advantage of TRAEN was even more pronounced
The results are show in the table below.
White cfu Blue cfu
STANDARD LIGATION: 3 2
TRAEN LIGATION 0,3 uL: 47 3
FIGURE 1 shows the outcome of the ligation experiment with (Left) and without (Right) the use of TRAEN.
 
Prokaryotic strains transformed successfully with higher efficacy are: E. coli, Dh5a, JM109, JM101
Lactococcus and Streptococcus, strains were not disclosed by customers.
(Get a discount on your purchase by adding a new strain or species)
The application of this new product is very simple.
Just add 0,3 ul of TRAEN to the DNA (10-100ng, see protocol) you want to transfer to E.Coli.
The rest of your protocol, which you worked hard to optimize , can be used as it is.
Protocols:
TRAEN Protocol
|
