Uneven PCR, Rapid method to walk unknown sequence, http://128.84.203.244
This paper describes a new polymerase chain reaction (PCR) cycling condition
to generate specific unknown fragments or genes directly from total DNA
instead of cloning genes from libraries.
For example, if you have a cDNA clone but missing the 5'end, or if you have a
gene coding sequence but you want to get the promoter region fast, or you
find that you can not get the damn Inversion PCR to work in your hands.
Before you decide to buy a kit to do these kind of work. Please try this
method first.
The essence of the method is to use two different annealing temperatures in
consecutive PCR cycles to
effectively amplify the target products while inhibiting the synthesis of non
specific products. We name this method "Uneven PCR". Under favorable
conditions, a desired DNA fragment or gene can be obtained and ready to clone
in just a few hours. The advantages of this method are simplicity, precision,
sensitivity, and speed. We believe that it will have a broad spectrum of
application in molecular biology.
This paper has been published in GENE 185(1997)195-197.
A similar method called TAIL PCR has also been published.
