siRNA / miRNA gene silencing Human COV-434

Get tips on using ChemiKine Brain Derived Neurotrophic Factor, Sandwich ELISA to perform ELISA Human - GDNF

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.

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

Get tips on using Anti-Collagen Type VII Antibody, clone 32,-VII to perform Immunohistochemistry Collagen VII [II-32] - Mouse Human -NA-

Get tips on using AllPrep DNA/RNA Micro Kit to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

Get tips on using EpiQuik Dnmt3A Assay Kit to perform DNA methylation profiling Whole genome profiling - MCF-7, MDA-MB-453 human breast cancer

Get tips on using Corning® Hepatocyte Culture Media Kit, 500 mL to perform 3D Cell Culture Media Human bladder cancer organoid

Get tips on using JetPrime to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)

Get tips on using Astrocyte Medium to perform Stem cell Differentiation media Glioma differentiation Human astrocytic lineage

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human airway epithelial cells