The RNA interference (RNAi) is used to inhibit gene expression or translation, by neutralizing targeted mRNA molecules. Two types of RNA molecules such as microRNA (miRNA) and small interfering RNA (siRNA) play a central role in RNAi. Few points have to considered to increase the transfection efficiency of siRNA. Always use healthy, actively dividing cells to maximize transfection efficiency. The confluency of cells should be between 50-70%. Always use the most appropriate siRNA concentration to avoid off-target effects and unwanted toxic side effects. Positive and negative controls should be used for each and every experiment to determine transfection efficiency.
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 Arp2 siRNA (h) to perform siRNA / miRNA gene silencing Human - T47-D Arp-2
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I would like to regulate the expression of a gene and in order to do that, I have purchased specific siRNA. After optimizing my transfection protocol and using electroporation I have achieved a 60-70% reduction of the gene of interest. However, I cannot observe a significant reduction of mRNA expression but only a reduction of protein. What might be the problem? Could the problem be in my cell treatment method?
Get tips on using Gβ 1 siRNA (h) to perform siRNA / miRNA gene silencing Human - T47-D GNB1
Get tips on using Stealth siRNA™ NOTCH2 to perform siRNA / miRNA gene silencing Human - THP-1 NOTCH2
Get tips on using Stealth siRNA™ NOTCH1 to perform siRNA / miRNA gene silencing Human - THP-1 NOTCH1
Get tips on using caveolin-1 siRNA (h) to perform siRNA / miRNA gene silencing Human - BEAS-2B CAV1
Get tips on using Flotillin-2 siRNA (h) to perform siRNA / miRNA gene silencing Human - BEAS-2B FLOT2
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