Protein quantification Yeast

Get tips on using MicroVue YKL-40 EIA to perform ELISA Human - Chitinase-3-Like Protein-1 (CHI3L1) or YKL-40

Get tips on using pYT379-CDK8-CycC-10xHis complex to perform Protein Expression Eukaryotic cells - S. frugiperda CDK8-CycC-10xHis complex

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 Mt-PA

Get tips on using pET30 Ek/LIC-Colα3(IV) NC1 to perform Protein Expression Prokaryotic cells - E. coli Colα3(IV) NC1

Get tips on using pcDNA™3.1 (+) Mammalian Expression Vector to perform Protein expression and purification Mammalian cells - CHO-K1 sRAGE

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - Human eutopic endometrial stromal cells

Get tips on using pFastBac1-A/reassortant/NYMC X-179-NP to perform Protein Expression Eukaryotic cells - S. frugiperda Influenza NP

Get tips on using HiTrap Q FF anion exchange chromatography column to perform Protein expression and purification Bacteria - Bacillus subtilis GCSF

Get tips on using Bac-to-Bac™ Baculovirus Expression System to perform Protein expression and purification Insect cells - Sf9 Drosha

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.