Feb 11, 2022

Public workspaceProtocol for Exo-CIP™ Rapid PCR Cleanup (#E1050) V.2

DOI
dx.doi.org/10.17504/protocols.io.bg9xjz7n
  • 1New England Biolabs
  • New England Biolabs (NEB)
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DOI: dx.doi.org/10.17504/protocols.io.bg9xjz7n
External link: https://neb.com/protocols/2019/01/16/protocol-for-exo-ciprapid-pcr-cleanup-e1050
Protocol Citation: New England Biolabs 2022. Protocol for Exo-CIP™ Rapid PCR Cleanup (#E1050). protocols.io https://dx.doi.org/10.17504/protocols.io.bg9xjz7nVersion created by New England Biolabs
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: June 08, 2020
Last Modified: February 11, 2022
Protocol Integer ID: 37911
Keywords: PCR, PCR Cleanup
Abstract
Exo-CIP™ Rapid PCR Cleanup Kit
  • Rapidly degrade residual PCR primers and dephosphorylate excess dNTPs after amplification
  • Reaction complete in 4 minutes
  • Thermolabile formulation can be heat inactivated in 1 minute at 80°C
  • PCR product can be used directly in downstream applications
  • Compatible with commonly-used reaction buffers
Materials
MATERIALS
ReagentExo-CIP Rapid PCR Cleanup Kit - 100 rxnsNew England BiolabsCatalog #E1050S
Safety warnings
Please see SDS (Safety Data Sheet) for hazards and safety warnings.
Transfer Amount5 µL PCR product to a new PCR tube and add Amount1 µL Exo-CIP A and Amount1 µL Exo-CIP B . The final volume is Amount7 µL .
Pipetting
Mix thoroughly and briefly centrifuge at Centrifigation1000 x g..
Centrifigation
Incubate the reaction tube for Duration00:04:00 at Temperature37 °C followed by Duration00:01:00 at Temperature80 °C .
Incubation
Submit Amount3 µL treated PCR product (in a range of 15-200 fmol) or less* for sequencing using BigDye™ Terminator v3.1 Cycle Sequencing Kit or store the treated samples at Temperature-20 °C for longer term storage.
Note
* A simple way to determine the amount of your amplicon is to load Amount3 µL on an agarose gel along with a known amount of a control DNA for comparison. Alternatively, direct measurement using fluorescent dye based kit (e.g., Qubit™) will ensure the proper amount of DNA is submitted.
AB
Size of PCR ampliconng of DNA (in 3 μl sample)
100 bp1 - 12
500 bp5 - 60
1000 bp10 - 120
3000 bp30 - 360
5000 bp50 - 600


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