protein-expression-and-purification-bacteria-escherichia-coli-p19

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Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

RNA RNA isolation / purification Tissue Human Blood / Serum / Plasma / Buffy coat

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

RNA RNA isolation / purification Tissue Mouse Blood / serum / plasma / buffy coat

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA

RNA RNA isolation / purification Tissue Rat Blood / Serum / Plasma / Buffy coat

Get tips on using Absolutely Total RNA Purification Kits to perform RNA isolation / purification Cells - primary rat cardiac fibroblasts

Products Agilent Technologies Absolutely Total RNA Purification Kits

Get tips on using Ni-NTA Superflow 96 BioRobot Kit (4) to perform Protein tag Purification of His-tagged proteins

Products Qiagen Ni-NTA Superflow 96 BioRobot Kit (4)

Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,

DNA DNA isolation / purification Cells Immortalized cell lines Renal cortical tubule epithelial cells

RNA RNA isolation / purification Tissue Human Colon

Get tips on using GeneJET RNA Purification Kit to perform AAA for reviews

Products Thermo Fisher Scientific GeneJET RNA Purification Kit

Get tips on using MinElute 96 UF PCR Purification Kit (24) to perform DNA isolation / purification Plasmid purification

Products Qiagen MinElute 96 UF PCR Purification Kit (24)

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.

RNA siRNA / miRNA gene silencing Mouse B16-BL6 p100/Nfkb2

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