Popular Applications for Specialized PCR: What to Know

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Specialized PCR applications include synthesizing double-stranded DNA from single-stranded templates, filling in recessed 3' ends, digesting protruding 3' overhangs, and preparing radioactive DNA probes. The Klenow fragment is highlighted for its ability to initiate synthesis from a primed single strand without the need for additional proteins, unlike DNA polymerase III, which requires single-stranded binding proteins. This makes Klenow a more straightforward option for certain applications. The discussion emphasizes the importance of choosing the right enzyme based on specific experimental needs. Understanding these distinctions can enhance the efficiency of PCR techniques in various genetic analyses.
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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.
 
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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
 
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Thank you Ian, that is just what I wanted to know! :smile: 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?
 
Klenow fragment can start synthesis from a primed single strand whereas DNA Pol III requires ssb protein in addition to a primed ss.
 

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