Tag:
Molecular Cloning
assay development
Restriction digest-PDF
General Information
Restriction digest involves the cutting of DNA at specific recognition sequences by enzymes.
General Procedure
Restriction Ligation in Genome Compiler
An intuitive cloning wizard for simulating...
assay development
Engineering BioBrick vectors from BioBrick parts/Dephosphorylation-PDF
To minimize self-ligated vector in your transformation, treat your linearized vector with a phosphatase to remove the 5' phosphates necessary for ligation. This should...
assay development
Engineering BioBrick vectors from BioBrick parts/DNA ligation-PDF
Materials
T4 DNA Ligase from NEB
10X T4 DNA Ligase buffer
Deionized, sterile H2O
Purified, linearized destination vector (in H2O)
Purified, linearized prefix part...
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
T4 Ligation-PDF
Overview
Standard DNA ligation using NEB T4 ligase to form a vector.
Materials
For a 10 μL ligation reaction:
3 μL sterile ddH2O
1 μL 10X ligation...
assay development
DNA ligation using NEB Quick Ligation Kit-PDF
Materials
We use the Quick Ligation Kit from NEB
Deionized, sterile H2O
Purified, linearized vector (in H2O)
Purified, linearized insert (in H2O)
Ligation Mix
This is...
gene therapy & gene editing
Cloning Protocol-PDF
PCR
There are two types of PCR polymerases used in the lab, TAQ and Phusion. Phusion is more accurate, having fewer mutations or errors in...
assay development
Restriction Digest: Partial-PDF
Abstract
This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes.
Materials
...