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PCR Based Moleclar Cloning

In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that it can be easily cloned into a plasmid of interest. You can use similar processes to add overhangs to your insert of interest for Gibson assembly. The steps following primer design and the PCR process itself are very similar to those outlined in our restriction cloning post with a few quirks specific to the PCR cloning process – please check out that post if you need a more detailed refresher on the downstream steps.

 

Standard Cloning PCR

a protocol which gives DNA that is clean enough for sequencing is the … This complicates cloning, as the supposedly blunt-ended PCR product often is not, …
www.mcb.uct.ac.za/pcrcond.htm

 

Addgene – Protocols: Basic cloning protocol

Steps for Cloning. Prepare DNA. See Recovering Plasmids from your Stab Culture protocol if you need to prepare DNA from a bacterial stock. See PCR protocol …
www.addgene.org/pgvec1?f=v&cmd=showfile&file=prot_cloning

 

In-Fusion™ Ready Vector Cloning Protocol-at-a-Glance
Cloning Protocol for In-Fusion™ PCR Cloning Kits. A. PCR Amplification for In-Fusion™ Ready Cloning. The sense and antisense primers that will be used to …
www.takara.co.kr/pds/shop/product/manual/pt3865-2.pdf

 

Purification of PCR Products in Preparation for Cloning

Selected Protocol from “Molecular Cloning: A Laboratory Manual”, Third Edition, Joseph Sambrook, David W. Russell. Purification of PCR Products in …
www.sigmaaldrich.com/…/Molecular_Biology/Key_Resources/Protocols/Purification_of_PCR_Products.html

 

Cloning PCR-Generated DNA Fragments

Protocol 2, PCR cloning, increases the yield and efficiency of blunt-ended cloning by incorporating a unit excess of restriction enzyme (compared to the …
wheat.pw.usda.gov/~lazo/methods/lazo/pcr-cloning.html –

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