DNA gel extraction / PCR product purification

Get tips on using pGEX-6P-1 Vector to perform Protein expression and purification Bacteria - Escherichia coli FleN

Get tips on using pQE-30 Xa Vector to perform Protein expression and purification Bacteria - Escherichia coli Siva1

Get tips on using ZR RNA MiniPrepTM kit to perform RNA isolation / purification Cells - primary human endothelial cells

Get tips on using mirVana® miRNA mimic to perform RNA isolation / purification Cells - primary human endothelial cells

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human CD14+ monocytes

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

Get tips on using RNeasy Fibrous Tissue Mini Kit to perform RNA isolation / purification Cells - primary mouse ventricles

Get tips on using RNeasy Fibrous Tissue Mini Kit to perform RNA isolation / purification Cells - primary mouse cardiomyocytes

Get tips on using QuantiFluor® dsDNA System to perform DNA quantification Colorectal aenocarenoma (SW48) - paraffin embeded

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.