siRNA / miRNA gene silencing Rat H19-7

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Rabbit eye retina/choroids

Get tips on using T-PER™ Tissue Protein Extraction Reagent to perform Protein isolation Tissue - Rabbit eye retina/choroids

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Get tips on using GenJet™ In Vitro DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines COS7

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 TRIzol Reagent to perform RNA isolation / purification Cells - immortalized ZR-75-1

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized DG-75

Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Human - SMMC-7721