Tag:
DNA amplification
gene therapy & gene editing
PNK Treatment of DNA Ends-PDF
Introduction
This protocol is used to add a phosphate group to the 5' end of a single or double stranded DNA molecule. Most primers, for...
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-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
TempliPhi-PDF
Materials
illustra TempliPhiâ„¢ 100/500 Amplification Kit
0.6mL PCR tubes
Procedure
Thaw sample buffer (red cap) and reaction buffer (blue cap) on ice.
Transfer 5 μL...
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...
assay development
Colony PCR-PDF
Protocol
Use a sterile toothpick or pipet tip to resuspend a plated colony in 50 μl sterile water.
store the colony resuspension at 4C...
Biologics (antibody+ADC)
Colony PCR-PDF
Zymolyase Solution:
Regular stock of zymolyase is 10 mg/ml diluted in water, mix by inverting, not all will go into solution (filter it) store...