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siRNA / RNAi /miRNA transfection Rat

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Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - Rat Retina

Products Thermo Fisher Scientific TRIzol Reagent

Get tips on using RNAsimple Total RNA Kit to perform RNA isolation / purification Tissue - Rat Placenta

Products Tiangen RNAsimple Total RNA Kit
hCas9 Product

Get tips on using hCas9 to perform CRISPR Rat - Activation CD2

Products Addgene hCas9

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 Magnetic mRNA Isolation Kit to perform RNA isolation / purification Tissue - Rat Cerebellum

Products New England BioLabs Magnetic mRNA Isolation Kit

Get tips on using Magnetic mRNA Isolation Kit to perform RNA isolation / purification Tissue - Rat Brainstem

Products New England BioLabs Magnetic mRNA Isolation Kit

Get tips on using Magnetic mRNA Isolation Kit to perform RNA isolation / purification Tissue - Rat Brain

Products New England BioLabs Magnetic mRNA Isolation Kit

Get tips on using SECISBP2 Polyclonal antibody to perform Immunohistochemistry Rat - SBP2

Products Proteintech Group SECISBP2 Polyclonal antibody

Get tips on using HighCell# ChIP kit to perform ChIP Rat - PCCL3

Products Diagenode HighCell# ChIP kit
EZ-ChIP™ Product

Get tips on using EZ-ChIP™ to perform ChIP Rat - NRK52E

Products Merck Millipore EZ-ChIP™

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