Silencer™ siRNA Construction Kit with manual
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Invitrogen™

Silencer™ siRNA Construction Kit with manual

The Ambion™ Silencer™ siRNA Construction Kit is for the synthesis of siRNAs by in vitro transcription of DNA templates andRead more
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Catalog NumberIncludes
AM1620Kit only
AM1620MKit with manual
Catalog number AM1620
Price (CNY)
16,968.00
Each
Add to cart
Includes:
Kit only
Price (CNY)
16,968.00
Each
Add to cart
The Ambion™ Silencer™ siRNA Construction Kit is for the synthesis of siRNAs by in vitro transcription of DNA templates and costs less per siRNA than chemical synthesis. The kit includes sufficient reagents to synthesize 15 siRNAs.
• Cut siRNA preparation time—generate up to 15 siRNAs in less than 24 hours
• Get hundreds of transfections per reaction
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Concentration2X
IncludesKit only
No. of Reactions15
Product LineSilencer™, Ambion™
Quantity15 reactions
Shipping ConditionDry Ice
Control TypeTranscription Control
Sample TypeIn vitro Transcribed siRNA
Unit SizeEach
Contents & Storage
DNA Hybridization Buffer, T7 Promoter Primer, Klenow Reaction Buffer, 10X dNTP, Exo– Klenow, T7 Enzyme Mix, T7 Reaction Buffer, 2X NTP Mix, Digestion Buffer, DNase, RNase, Sense Control DNA, and Antisense Control DNA should be stored at –20°C. siRNA Binding Buffer, siRNA Wash Buffer, Filter Cartridges, and Tubes should be stored at room temperature. Nuclease-free Water may be stored at any temperature.

Frequently asked questions (FAQs)

What are the benefits of using a vector to deliver RNAi?

Vector technologies allow you to:

Achieve transient or stable target knockdown
Perform RNAi in any cell type, even hard-to-transfect, primary, and non-dividing cells
Regulate gene inhibition with inducible siRNA expression
Select for a pure population of cells stably expressing an siRNA sequence
Control gene expression in vivo with tissue-specific promoters

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

Why are my cells dying after transfection?

We would suggest running a transfection reagent control only to determine if your cells are sensitive to the transfection reagent. Additionally, you can try using different cell densities and siRNA concentrations to diminish any toxic effects from the transfection itself.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I transfected my siRNA and the mRNA levels are down, but the protein is not. Why is that?

In some cases, knockdown of a protein can be affected by other variables such as protein turnover rate, even though the RNA is knocked down. Additionally, a longer time course may be needed to see an effect on protein compared to mRNA.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I am not getting my target knockdown. What could be the cause of this?

Please see the following possibilities and suggestions:

- How many siRNA did you test? Is there any knockdown? If there is no knockdown (<10%) in any of the siRNA, then the assay is likely the problem. Try using a different qRT-PCR assay to assess knockdown.
- What was the positioning of the qRT-PCR assay target site relative to the cut site for the siRNA? If greater than 3,000 bases away, the problem could be alternative splice transcripts.
- What are the Cts for the experiment? They should be below 35 in a 40-cycle qRT-PCR experiment.
- Did you confirm the siRNA got into the cell? We recommend using a validated positive control siRNA to check the transfection efficiency.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I am not getting any knockdown with my siRNA. What do you suggest I try?

Please see the following possibilities and suggestions:

- Were the mRNA levels checked? The most reliable method is real-time PCR. In some cases, knockdown of a protein can be affected by other variables, such as protein turnover rate, even though the RNA is knocked down.
- How is the RNA being isolated? Has the quality of the isolated RNA been checked? Ensure that the RNA has not been degraded.
- Was a positive control used? This can help to determine whether the reagents are working and whether the siRNA was delivered correctly to the cell. Run your experiment in parallel with the positive control siRNA.
- Was a transfection control used? What is the percentage of transfected cells?
- Was a time course used? Generally, gene silencing can be assessed as early as 24 hours posttransfection. However, the duration and level of knockdown are dependent on cell type and concentration of siRNA.
- Was optimization of transfection conditions performed? You can try using different cell densities and siRNA concentrations.
- Which concentration of siRNA did you use? We recommend testing multiple concentrations between 5 nM and 100 nM.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.