Klenow Fragment

[rockind78]

I know what it is, but my question is: What are some (specifically) popular applications? I am guessing some sort of specialized PCR??? Any thoughts would be appreciated.

[iansmith]

I know we use it to double strand DNA for electrophoretic mobility shift assay (EMSA) and DNA footprintings.

From the following site
http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/enzymes/klenow.html

1. Synthesis of double-stranded DNA from single-stranded templates
2. Filling in recessed 3' ends of DNA fragments
3. Digesting away protruding 3' overhangs
4. Preparation of radioactive DNA probes

[rockind78]

Thank you Ian, that is just what I wanted to know!! [:)] This brings me to my next question I guess, being, what advantage would there be in utilizing JUST klenow fragment in synthesis of DS DNA fragments from SS DNA versus DNA polymerase III?

[iansmith]

Klenow fragment can start synthesis from a primed single strand whereas DNA Pol III requires ssb protein in addition to a primed ss.