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Get tips on using Maxwell® 16 LEV simplyRNA Purification Kit to perform RNA isolation / purification Tissue - Human Muscles

Products Promega Maxwell® 16 LEV simplyRNA Purification Kit

Get tips on using PE Mouse Anti-Human CD26 Clone L272 to perform Flow cytometry Anti-bodies Human - CD26

Products BD Biosciences PE Mouse Anti-Human CD26 Clone L272

Get tips on using PE Mouse Anti-Human CD30 Clone BerH8 to perform Flow cytometry Anti-bodies Human - CD30

Products BD Biosciences PE Mouse Anti-Human CD30 Clone BerH8

Get tips on using Monoclonal Mouse Anti-Human Cytokeratin, Clone MNF116 to perform Flow cytometry Anti-bodies Human - Keratin

Products Agilent Technologies Monoclonal Mouse Anti-Human Cytokeratin, Clone MNF116

Get tips on using A2B5 Antibody, anti-human/mouse/rat, APC to perform Flow cytometry Anti-bodies Human - A2B5

Products Miltenyibiotec A2B5 Antibody, anti-human/mouse/rat, APC

Get tips on using Monoclonal Anti-Laminin antibody produced in mouse to perform Western blotting Laminin subunit Beta-2

Products Sigma-Aldrich Monoclonal Anti-Laminin antibody produced in mouse

Get tips on using Monoclonal Anti-ATG5 antibody produced in mouse to perform Autophagy assay cell type - CaCo-2

Products Sigma-Aldrich Monoclonal Anti-ATG5 antibody produced in mouse

Get tips on using Monoclonal Anti-ATG12 antibody produced in mouse to perform Autophagy assay cell type - CaCo-2

Products Sigma-Aldrich Monoclonal Anti-ATG12 antibody produced in mouse

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.

DNA Plasmid Isolation Shiga toxin-producing E. coli

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.

DNA Plasmid Isolation E. coli-S. cerevisiae transconjugate

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