May 16, 2020
Version 1
Enzymatic PCR Cleanup Protocol (NEB #M0525) V.1
- New England Biolabs1
- 1New England Biolabs
- New England Biolabs (NEB)Tech. support phone: +1(800)632-7799 email: [email protected]
Protocol Citation: New England Biolabs 2020. Enzymatic PCR Cleanup Protocol (NEB #M0525). protocols.io https://dx.doi.org/10.17504/protocols.io.7r4hm8w
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 28, 2019
Last Modified: March 30, 2021
Protocol Integer ID: 28188
Keywords: exo sap, exo-sap, exosap, exocip, exo-cip, exo-cip, PCR Cleanup, PCR Clean up, enzymatic pcr cleanup protocol, intestinal alkaline phosphatase, need for multiple phosphatase, snp analysis recombinant for purity, snp analysis recombinant, multiple phosphatase, native enzyme faster reaction setup, less enzyme, active in any restriction enzyme buffer, quick cip, pcr product, enzyme, restriction enzyme buffer, dna sequencing, shorter incubation time flexible reaction condition, cip, shorter incubation time, phosphates from dna
Abstract
Enzymatic PCR Cleanup Protocol (NEB #M0525) uses Quick CIP; Quick CIP is a heat-labile version of calf intestinal alkaline phosphatase (CIP) purified from a recombinant source.
Features
- Rapid and irreversible heat inactivation eliminates unwanted activity
- Improved storage stability versus native enzyme
- Faster reaction setup (no supplemental additives like zinc required) and shorter incubation time
- Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
- Less enzyme required (high specific activity), resulting in a lower cost per reaction
- No need for multiple phosphatases (Quick CIP removes 5′- and 3′- phosphates from DNA, RNA and dNTPs )
- Active on unincorporated dNTPs in PCR products - improves DNA sequencing and SNP analysis
- Recombinant for purity, consistency and value
Materials
MATERIALS
Quick CIP New England BiolabsCatalog #M0525
Exonuclease I (E. coli)New England BiolabsCatalog #M0293
Shrimp Alkaline Phosphatase (rSAP)New England BiolabsCatalog #M0371
Troubleshooting
Safety warnings
Please see SDS (Safety Data Sheet) for hazards and safety warnings.
Note
Note that 0.5 μl of Exo I (NEB #M0293) and 1 μl of rSAP (NEB #M0371) can be used in the same volume.
Incubate the mix at the 37 °C for 00:15:00 .
Inactivate both enzymes at 80 °C for 00:15:00 .
PCR products are ready for downstream application.