BAA10@phoenix.cambridge.ac.uk (04/13/89)
We have had success only after carefully purifying the products. Our procedure is; gel purify, phenol extract, chloroform extract, ethanol precipitate, redissolve and quantitate, end fill, kinase. We then titrate ligations over a wide range of vector:insert ratios and usually get recombinants. You can also engineer sticky ends to your products by adding the requisite sites to your primers. Ligation is then more efficient. This would be the method of choice for amplifying from a well characterized template, but we have had some peculiar results when trying to clone this way from genomic DNA. /*