DNA methylation profiling Gene specific profiling whole blood (human)

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 Gibco™DMEM, low glucose, pyruvate to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Get tips on using Mesenchymal Stem Cell Osteogenic Differentiation Medium to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Get tips on using Q5® Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Human - Point mutation LNCaP Androgen Receptor splice variant (AR-V)

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

Get tips on using Gibco™ MEM α, GlutaMAX™ Supplement, no nucleosides to perform Stem cell Differentiation media Human oogonial stem cells differentiation into oocytes

Get tips on using DMEM/Ham's F-12 liquid medium w/o L-Glutamine to perform Stem cell culture media Human Tendon Stem/Pluripotence cells (TSPCs)

Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.