Tag:
PCR
gene therapy & gene editing
Site-directed mutagenesis-PDF
General Information
Site-directed mutagenesis can be used to change particular base pairs in a piece of DNA. There are several methods for achieving this. The...
assay development
PCR Overlap Extension-PDF
Overview
Create long DNA fragments from shorter ones. This method is also called "Splicing by Overlap Extension" or SOEing.
Procedure
Design Primers:
These primers are like...
assay development
PCR techniques-PDF
Several techniques have have been derived from the basic polymerase chain reaction (PCR). Below is an overiew of important PCR methods with links to...
assay development
PCR-PDF
Overview
Standard polymerase chain reaction (PCR) protocol for amplification of DNA.
Materials
For a 50 μL PCR reaction:
35 μL H2O
5 μL 10X PCR buffer
5...
assay development
Annealing and primer extension with Taq polymerase-PDF
This protocol uses annealing and primer extension to generate a short fragment of DNA (~100 bp) using Taq polymerase. The DNA fragment can be...
gene therapy & gene editing
Stitching Genes by PCR-PDF
Introduction
This method allows you to "stitch" genes or coding sequences together when there are no convenient restriction sites at the junction point. It is...
assay development
Whole Plasmid PCR-PDF
Overview
It is imperative to use 5'-phosphorylated primers if the nicked DNA is going to be repaired downstream with ligase. PCR should be limited to...
assay development
Engineering BioBrick vectors from BioBrick parts/Colony PCR-PDF
Materials
PCR SuperMix High Fidelity
VF2 primer (5'-TGCCACCTGACGTCTAAGAA-3')
VR primer (5'-ATTACCGCCTTTGAGTGAGC-3')
Deionized, sterile H2O
strip of PCR tubes
2-log DNA ladder
0.8% E-Gel® from...