Apr 25, 2022

Public workspacePCR mixture and condition (2X SUPERGREEN PCR MASTER MIX) V.3

This protocol is a draft, published without a DOI.
  • 1Kaohsiung Medical University
Yin-Tse Huang
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Protocol Citation: Yin-Tse Huang 2022. PCR mixture and condition (2X SUPERGREEN PCR MASTER MIX). protocols.io https://protocols.io/view/pcr-mixture-and-condition-2x-supergreen-pcr-master-b755rq86Version created by Yin-Tse Huang
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: In development
We are still developing and optimizing this protocol
Created: April 25, 2022
Last Modified: September 13, 2023
Protocol Integer ID: 61341
Abstract
PCR mixture and condition (2X SUPERGREEN PCR MASTER MIX)
For generic primers
For generic primers
PCR mixture PCR condition for generic primers
ABC
ComponentVolumeFinal conc.
Template (< 1 µg)X µL-
Forward Primer (10 µM)1 µL0.4 µM
Reverse Primer (10 µM)1 µL0.4 µM
2x Master Mix12.5 µL-
ddH20up to 25 µL-
Total volume25 µL-

PCR condition for generic primers

ABCD
StepTempSecCycle
Initial denaturation95 ºC180
Denaturation95 ºC3035 cycles
Annealing53-58 ºC30
Extension72 ºC60
Final extension72 ºC210
Preservation4 ºC∞

For barcoded primers
For barcoded primers
PCR mixture for barcoded primers

ABC
ComponentVolumeFinal conc.
Template (< 1 µg)X µL-
Forward Primer (10 µM)5 µL2 µM
Reverse Primer (10 µM)5 µL2 µM
2x Master Mix12.5 µL-
ddH20up to 25 µL-
Total volume25 µL-

PCR condition for barcoded primers

ABCD
StepTempSecCycle
Initial denaturation95 ºC180
Denaturation98 ºC2035 cycles
Annealing64 ºC15
Extension72 ºC210
Final extension72 ºC210
PreservationPreservation4 ºC∞