Protein isolation Tissue

Get tips on using Gentra Puregene Mouse Tail Kit (4 g) to perform DNA isolation / purification Tissue - murine tail biopsies

Get tips on using ARCTURUS® PicoPure® DNA Extraction Kit to perform DNA isolation / purification Tissue - murine tail biopsies

Get tips on using Easy-Spin (DNA free) Total RNA Extraction Kit to perform RNA isolation / purification Tissue - Mouse Cornea

Get tips on using VWR Life Science RiboZol™ RNA Extraction Reagent to perform RNA isolation / purification Tissue - Human Gallbladder

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.

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 RIPA Buffer to perform Protein isolation Mammalian cells - Human lung fibroblasts

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Mouse Epididymal fat

Get tips on using TRI Reagent® to perform Protein isolation Mammalian cells - Mouse_Brown fat

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - Caco-2