Please explain to me how all these work, and work together better then allready

Question

Please explain to me how all these work, and work together better then allready done?!?

•       Exonuclease I E. coli: Removes oligonucleotides post PCR

•       T7 Exonuclease:

•       DNA Polymerase I large (Klenow) fragment: retains polymerization fidelity of holoenzyme w/o degrading 5’ termini

•       Alkaline Phosphatase: catalyze 5’ phosphate removal from DNA, RNA, ribo-deoxyrebonucleoside triphosphate

•       SAPas (shrimp)

•       CIPase (calf intestine)

•       T4 DNA Ligase: Catalyzes formation of phosphodiester bond between 5’-phosphate and 3’ hydroxyl termini in duplex DNA or RNA

Explanation / Answer

Exonuclease I E. coli: Removes oligonucleotides post PCR

This enzyme is a monomer catalyses the removal of nucleotides from single stranded DNA in the 3' to 5' direction. with free hydroxyl group at 3' and releases mono or dinucleotides. It is inhibited by 3' terminal phosphate. It cannot digest double stranded DNA.It produces 5'phosphate mononucleotides from 3' hydroxyl terminus. IT removes left over primers after PCR

•      T7 Exonuclease: acts in 5' to 3' direction catalysing the removal of 5' mononucleotides from duplex DNA. It degrades 5' phosphorylated or 5' dephosphorylated DNA.

•       DNA Polymerase I large (Klenow) fragment: retains polymerization fidelity of holoenzyme w/o degrading 5’ termini

•       Alkaline Phosphatase: catalyze 5’ phosphate removal from DNA, RNA, ribo-deoxyrebonucleoside triphosphate

•       SAPas (shrimp): Shrimp Alkaline phosphatase removes 5' phosphatase from 5' and 3' ends of DNA and RNA. It removes excess dNTP's after PCR

•       CIPase (calf intestine)

•       T4 DNA Ligase: Catalyzes formation of phosphodiester bond between 5’-phosphate and 3’ hydroxyl termini in duplex DNA or RNA

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