Immunohistochemistry Anti-rabbit IgG Donkey Rabbit

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat articular chondrocytes

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat articular chondrocytes

Get tips on using JetFlex™ Genomic DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram positive Actinomycytes

Get tips on using GenElute™ Bacterial Genomic DNA Kit to perform DNA isolation / purification Bacteria - Gram positive Actinomycytes

Get tips on using HiPurA™ Streptomyces DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram positive Actinomycytes

Get tips on using E.Z.N.A.® Plasmid Mini Kit I, (Q-spin) to perform Plasmid Isolation Actinomyces odontolyticus

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Get tips on using GeneArt™ CRISPR Nuclease Vector with OFP Reporter Kit to perform CRISPR Human - Activation CD20

Get tips on using GeneArt™ CRISPR Nuclease Vector with OFP Reporter Kit to perform CRISPR Human - Activation ERBB2

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.