rna-isolation-purification-cells-primary-mouse-dorsal-root-ganglion-neurons

Get tips on using pMIR-REPORT™ miRNA Expression Reporter Vector System to perform Reporter gene assay luciferase - HEK 293 human embryonic kidney cells

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality hot-start DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction

Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells

Get tips on using in situ Cell Death Detection Kit, POD to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells

Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - BHK-21

A restriction enzyme or restriction endonuclease is defined as a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at or near that site, known as restriction site or target sequence. The four most common types of restriction enzymes include: Type I (cleaves at sites remote from a recognition site), Type II (cleaves within or at short specific distances from a recognition site), Type III (cleave at sites a short distance from a recognition site), and Type IV (targets modified DNA- methylated, hydroxymethylated and glucosyl-hydroxymethylated DNA). The most common challenges with restriction digest include- 1. inactivation of the enzyme, 2. incomplete or no digestion, and 3. unexpected cleavage. The enzyme should always be stored at -20C and multiple freeze-thaw cycles should be avoided in order to maintain optimal activity. Always use a control DNA digestion with the enzyme to ensure adequate activity (to avoid interference due to high glycerol in the enzyme). For complete digestion, make sure that the enzyme volume is 1/10th of the total reaction volume, the optimal temperature is constantly maintained throughout the reaction, the total reaction time is appropriately calculated based on the amount of DNA to be digested, appropriate buffers should be used to ensure maximal enzymatic activity, and in case of a double digest, make sure that the two restriction sites are far enough so that the activity of one enzyme cannot interfere with the activity of the other. Star activity (or off-target cleavage) and incomplete cleavage are potential challenges which may occur due to suboptimal enzymatic conditions or inappropriate enzyme storage. To avoid these, follow the recommended guidelines for storage and reactions, and always check for the efficacy of digestion along with purification of digested products on an agarose gel.

Get tips on using Gibco™DMEM/F-12, no glutamine to perform Stem cell Differentiation media hPSCs or iPSCs differentiation into Lung progenitor cells