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Description

We can label the ends of siRNA with a variety of fluorescent markers. The fluorescently labeled siRNA could be observed by flow cytometry, fluorescence microscope, and laser confocal microscope to determine the transfection efficiency and optimize the transfection conditions. The fluorescently labeled siRNA can also be used as the intracellular localization of siRNA and double labeling experiment (with labeled antibody) to track the siRNA transfected cells during the transfection process, and combine the transfection with the down-regulation of target protein expression.

The 5'-end marker of the antisense chain will affect its gene silencing activity, so it is not recommended to mark at this site. The terminal modifications on the other three sites have little effect on the silencing activity. We recommend modifying the 5'-end of the sense chain, which is recognized as the best chemical marker so far.

Result Analysis

For RNAi experiments, we usually need to know that the expression of a specific gene in a specific cell before and after siRNA is introduced, in order to judge whether siRNA plays a role in gene knockdown. The above judgment can be achieved by qPCR. There are mainly two ways: One is to determine the change of mRNA quantity of a specific gene before and after siRNA introduction, that is, absolute quantification. The other is to detect the relative expression changes of a specific gene and a housekeeping gene before and after siRNA introduction, that is, relative quantification. Generally, the gene knockdown effect of siRNA is detected by relative quantification.

Note

• The siRNA provided by us is all purified by HPLC. The leading time is around 9 working days.
• Our siRNA directly acts on the mRNA of the target gene. Therefore, we recommend conducting qPCR after receiving our products to determine the change of mRNA expression level of the target gene, and then confirm whether the siRNA synthesized by us is effective.
• For larger specifications (>100 OD), please email us your requirement at tech@sbsbio.com.

Instruction: Protocol

Only for research and not intended for treatment of humans or animals

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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory