siRNA / miRNA gene silencing Human NCL-H40

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.

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.

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase

Get tips on using siRNA Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - IEC Cationic lipid based

Get tips on using AChE shRNA Plasmids (h) to perform shRNA gene silencing Human - TF‐1 AChE

Get tips on using MCM4 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - SiHa MCM4

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Get tips on using pSilencer 2.1-U6 Hygro to perform shRNA gene silencing Human - SHSY5Y Beclin 1

Get tips on using siRNA Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - IEC-6 Cationic lipid based

Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10