PCR (Q5 Polymerase)

This is for a 50 µl reaction.

Contents

1. Reaction Setup
2. PCR Reaction

1. Reaction Setup

Component	Amount
5X Q5 Reaction Buffer	10 µl
10 mM dNTPs	1 µl
10 µM Forward Primer	2.5 µl
10 µM Reverse Primer	2.5 µl
Template DNA	variable (< 1,000 ng total)
Q5 Polymerase	0.5 µl
Nuclease-Free Water	to 50 µl

2. PCR Reaction

  Initial Denaturation    98°C    30 seconds
  25–35 Cycles
      98°C 5–10 seconds
      50–72°C 10–30 seconds
      72°C 20–30 seconds/kb
  Final Extension 72°C 2 minutes
  Hold 4–10°C