Troubleshooting For Molecular Cloning

Posted by Bethany Peacock on November 07, 2019

Troubleshooting For Molecular Cloning

Troubleshooting Guide For Cloning. Troubleshooting Guide for Cloning Incubate plates at lower temperature (25 – 30°C). Transformation may need to be carried out using a strain that exerts tighter transcriptional control over the DNA

PCR Troubleshooting Guide. PCR troubleshooting guide. In molecular cloning, after the synthesis of cDNA from mRNA molecule templates, a PCR program must be designed to amplify the gene of interest, as well as add additional elements such as restriction sites or detection/purification tags. Intrinsic properties of gene sequences such as high GC content, long stretches of the same polynucleotide, and sequences encoding hairpin loop structures can all hinder PCR efficiency.

DNA Library Protocols. DNA library preparation. If the goal is to create a genomic DNA library, this first step is to extract genomic DNA (please see the protocol for DNA extraction ).Whereas the first step to generate a cDNA library relies on mRNA extraction (please see the protocol for RNA extraction ).Afterward, the mRNA is converted to cDNA through the catalytic activity of reverse transcriptase enzyme (please

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