Product Introduction
This enzyme has exonuclease activity that degrades single-stranded DNA from the 3 '-5' direction, progressively releasing DNA 5' monophosphate and leaving intact 5' terminal dinucleotides. The enzyme was derived from E.coli Exoi gene plasmid and purified by recombinant expression. This enzyme is mostly used to degrade digestion primers after PCR amplification. This enzyme is not active on double-stranded DNA and 3 'OH terminal DNA strands that are blocked by phosphoryl or acetyl groups.
Application
1. Remove primers from PCR mixture
2.PCR products were sequenced prior to PCR mutagenesis using large primers in a single tube
3. Remove the single strand of DNA containing the 3' hydroxyl terminus from the nucleic acid mixture
4. Detect the presence of a single strand of DNA containing the 3' hydroxyl terminus
Precautions
(1) 1×ExoIBuffer: 67mMGlycine-KOHpH9.5, 6.7mMMgCl2, 10mm2-mercaptoethanol. This enzyme is also active in conventional Pcrbuffers.
(2) The optimum reaction temperature of this enzyme is 37℃, and it can be inactivated at 80℃for 20min.
(3) The enzyme cannot cut double-stranded DNA, so single-stranded DNA containing secondary structures needs to be denatured before it can be fully digested.
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