DNA methylation profiling Gene specific profiling UMR-106

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 QIAamp PowerFecal Pro DNA Kit (50) to perform DNA isolation / purification Tissue - fecal sample

Get tips on using QIAamp DSP DNA Blood Mini Kit to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using EZ1 DSP DNA Blood Kit (48) to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using QIAamp DNA Blood Mini QIAcube Kit to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using Phusion Hot Start II DNA Polymerase to perform PCR Hot start PCR - Bacterial DNA

Get tips on using HiPurA™ Streptomyces DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram positive Actinomycytes

miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

Get tips on using mericon DNA Bacteria Plus Kit (50) to perform DNA isolation / purification Bacteria - Gram positive Clostridium botulinum