DNA methylation profiling

Get tips on using MaXtract High Density (100 x 15 ml) to perform DNA isolation / purification Tissue - small intestine

Get tips on using Hot StarTaq Master Mix Kit (2500 U) to perform PCR Hot start PCR - Mammalian DNA

Get tips on using QuantiTect SYBR Green RT-PCR Kit (1000) to perform PCR Conventional / Qualitative PCR - mammalian DNA

Get tips on using Quant-iT™ PicoGreen® dsDNA Assay Kit to perform DNA quantification Mouse - NIH 3T3

Get tips on using Quant-iT™ PicoGreen® dsDNA Assay Kit to perform DNA quantification Human - Hep G2

Get tips on using Quant-iT™ PicoGreen® dsDNA Assay Kit to perform DNA quantification Human - SH-SY5Y

Get tips on using illustra tissue and cells genomicPrep Mini Spin Kit to perform DNA isolation / purification Tissue - kidney

Get tips on using GenLadder 50bp (ready-to-use) with dye Orange G to perform DNA Ladder 50 bp

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.

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.