cloning question - (Jun/24/2001 )
I have worked in molecular biology for 8 months now and found the same thing. The classical purifications techniques such as gel extraction and ethanol precipitation seem to give low yields when carried more than once in a typical cloning experiment. One way to overcome these difficulties is to purchase commercial kits. Silica based kits such as Geneclean from Bio101 give much better yields, I have found. In addition, the proceedure is faster and eliminates the need for phenol/chloroform extractions. What the molecular cloning manual does not tell you is how exactly to carry out a complete cloning experiment. For example, how to incorporate restriction sites into primers, a very popular cloning choice, and the various downfalls of such a method.If you want more details of how I clone (although I'm no expert) please e-mail
I use cloning methods on Molecular Cloning Lab Mannual(second edition) ,but always get very low result.What's the reason?
Do you have any specific problems or is it just cloning in general?If notcleaning up rxns with colomns is a useful procedure to get reasonable quality DNA but other than that try playing with the experiments, you dont HAVE to follow SOME protocols word for word, try and find what works for you and stick to it!