rna-isolation-purification-tissue-rat-spinal-cord

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Get tips on using Quick Amp Labeling Kit-one color to perform Microarray RNA amplification & Labeling - Mouse Myofibers Cy3- or/and Cy5

Products Agilent Technologies Quick Amp Labeling Kit-one color

Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)

Products Santa Cruz Biotechnology Rock-2 siRNA and shRNA Plasmids (h)

Get tips on using Color-coded Prestained Protein Marker, Low Range (1.7-42 kDa) #13070 to perform Protein Ladder Prestained

Products Cell Signaling Technology Color-coded Prestained Protein Marker, Low Range (1.7-42 kDa) #13070

Get tips on using Color-coded Prestained Protein Marker, High Range (43-315 kDa) #12949 to perform Protein Ladder Prestained

Products Cell Signaling Technology Color-coded Prestained Protein Marker, High Range (43-315 kDa) #12949

Get tips on using Color-coded Prestained Protein Marker, Broad Range (10-250 kDa) #74124 to perform Protein Ladder Prestained

Products Cell Signaling Technology Color-coded Prestained Protein Marker, Broad Range (10-250 kDa) #74124

Short hairpin or small hairpin RNA (shRNA) is artificial RNA, which has a hairpin loop structure, and uses inherent microRNA (miRNA) machinery to silence target gene expression. This is called RNA interference (RNAi). These can be delivered via plasmids or viral/bacterial vectors. Challenges in shRNA-mediated gene silencing include: 1. Off-target silencing, 2. Packaging shRNA encoding lentivirus, and 3. Stable transduction in cells. RNAi have been designed to have anywhere from 19-27 bs, but the most effective design has 19 bp. In case commercial shRNAs are not available, potential target sites can be chosen within exon, 5’- or 3’ UTR, depending on which splice variants of the gene are desired. One should use the latest algorithms and choose at least two different sequences, targeting different regions, in order to have confidence in overcoming off-target effects. A BLAST search after selecting potential design will eliminate potential off-target sequences. For the second challenge, sequencing the vector using primers for either strand (50-100 bp upstream) is suggested, along with using enzymatic digestion on agarose gel for the vector. Next, once the shRNA-containing vector is packaged in a virus, it is important to check the viral titer before transduction. Finally, using a marker in the lentiviral vector (fluorescent protein or antibiotic resistance), along with qPCR for target gene expression can help in determining efficacy of transduction and shRNA on its target site.

RNA shRNA gene silencing Mouse Prostate cancer cell lines (DU145 and PC3) CD24 lentiviral particles

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray Human - Precision cut lung slices Target preparation kit (RNA Amplification + Hybridization + control)

Products Thermo Fisher Scientific GeneChip® HT 3' IVT PLUS Reagent Kit

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Cellular assays ROS assay cell type mouse splenocytes

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Cellular assays ROS assay cell type HeLa

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Cellular assays ROS assay cell type MCF-7

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