Site Directed Mutagenesis (SDM) Rat Point mutation H9C2

Get tips on using ON-TARGETplus Rat Fyn siRNA to perform siRNA / miRNA gene silencing Rat - Schwann cells Fyn

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 has 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 the efficacy of transduction and shRNA on its target site.

Get tips on using ON-TARGETplus Rat Tp53 (24842) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - C6 (rat glioma) p53

Get tips on using Rat VEGF ELISA Kit (ab100786) to perform ELISA Rat - VEGF

Get tips on using Rat VEGF Quantikine ELISA Kit to perform ELISA Rat - VEGF

Get tips on using Rat prolactin,PRL ELISA Kit to perform ELISA Rat - PRL

Get tips on using Rat GDNF ELISA Kit (ab213901) to perform ELISA Rat - GDNF

Get tips on using Rat Fibronectin ELISA Kit (ab108850) to perform ELISA Rat - Fibronectin

Get tips on using Mouse Rat Estradiol ELISA Kit to perform ELISA Rat - Estradiol

Get tips on using Rat CX3CL1/Fractalkine DuoSet ELISA to perform ELISA Rat - CX3CL1