Prerequisite Protocols

• PCR Primer Design

Materials

• Q5 High-Fidelity DNA Polymerase | NEB
• Q5 Reaction Buffer (already comes with Q5 Polymerase order)
• Deoxynucleotide (dNTP) Solution Mix | NEB
• Forward PCR Primer Working Stock 10uM
• Reverse PCR Primer Working Stock 10uM
• Sterile Distilled Water
• Template DNA (miniprep, genomic isolate, etc)
• Sterile 0.2mL PCR Tubes
• Sterile 1.5mL Tubes
• Pipette (2.5uL, 20uL, 200uL)
• Purple Loading Dye 6x or your favorite loading dye

Equipment

• PCR Machine
• Thermoshaker set to 37C 1000rpm
• Vortexer
• Ice Bucket with Ice or Cooling Block for 0.2mL and 1.5mL Tubes

Step By Step