I will be performing binding studies of various zinc fingers to DNA using ITC, which unfortunately requires a large amount of material. Commercially synthesized DNA would cost roughly US$10-$20 per experiment for 16-36 bp sequences.
To save money, I have considered cloning many copies (100-300) of the DNA binding sites of interest into a high copy plasmid such as pUC19, each copy flanked by restriction sites. Then I would grow bacterial cultures, isolate plasmid at large scale, and isolate the DNA sequences using restriction enzyme digestion followed by mung bean nuclease digestion, then polyacrylamide gel electrophoresis and band purification.
Is this a reasonable strategy? How do other researchers go about expressing micromolar concentrations of 12-40 bp DNA fragments?
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