siRNA / miRNA gene silencing Rat Cardiomyocyte (H9C2) HIF-1α

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. The resulting amplicons are generally detected by gel electrophoresis and for some further applications like cloning, sequencing, amplicon product needs to be recovered from the gel and subsequently purified. However, non-specific product amplification and primer-dimer formation during set-up make gel extraction difficult. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

Get tips on using Gibco™ DMEM, high glucose to perform Mammalian cell culture media RAOEC

Get tips on using Anti-acetyl-Histone H3 Antibody to perform ChIP acH3 - Rabbit Human YFP

Get tips on using Atg7 (D12B11) Rabbit mAb to perform Autophagy assay cell type - Hippocampal neural stem cells

Get tips on using Anti-acetyl-Histone H4 Antibody to perform ChIP acH4 - Rabbit Sheep YFP Tag

Get tips on using Anti-p62/SQSTM1 antibody produced in rabbit to perform Autophagy assay cell type - Hippocampal neural stem cells

Get tips on using Kapa Biosystems HIFI HOTSTART READY MIX to perform PCR Hot start PCR - Bacterial DNA

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

Get tips on using GenJet™ In Vitro DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)

Get tips on using Rapid Sequencing Kit to perform Whole Genome Amplification Human