CRISPR Rat Deletion INS-1 832/13

Get tips on using IRIS9 Prestained Protein Ladder（15 to 180 kDa） to perform Protein Ladder Prestained

Get tips on using Anti-Ctip1/BCL-11A antibody [14B5] (ab19487) to perform ChIP Anti-bodies CtIP/BCL11A

Get tips on using Blu11 (BlueAQUA) Prestained Protein Ladder（10 to 180 kDa） to perform Protein Ladder Prestained

Get tips on using Blu10 (BlueRAY) Prestained Protein Ladder（10 to 180 kDa） to perform Protein Ladder Prestained

Get tips on using PageRuler™ Prestained Protein Ladder, 10 to 180 kDa to perform Protein Ladder Prestained

Get tips on using Iscove’s Modified Dulbecco’s Medium to perform Stem cell Differentiation media iPSCs or hESCs differentiation into cerebellar neuroepithelium (NE)

Get tips on using DMEM/F-12 with 15 mM HEPES to perform Stem cell culture media hESC lines H9, H1

Get tips on using CD137 (4-1BB) Monoclonal Antibody (17B5), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD137

Get tips on using CD137 (4-1BB) Monoclonal Antibody (17B5), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD137

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.