Western blot 1,4 β-N-acetyl-D-glucosamine

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

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 BLOCK-iT™ Adenoviral RNAi Expression System, pAd/BLOCK-iT™-DEST RNAi Gateway Vector to perform shRNA gene silencing Mouse - P19 Foxm1

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Get tips on using QIAamp DNA Blood Midi Kit 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

When extracting nucleic acids from cell cultures. thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

Get tips on using EZ1 DNA Blood 200 µl Kit (48) to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using QIAamp DNA Blood BioRobot MDx Kit (12) to perform DNA isolation / purification Tissue - blood / plasma