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Custom RNA, Stealth, HPLC - FAQs

View additional product information for Custom RNA, Stealth, HPLC - FAQs (100001864)

13 product FAQs found

What controls should I use for my in vivo siRNA studies?

We offer the following in vivo siRNA controls:

Invitrogen In Vivo Negative Control #1 siRNA (Cat. No. 4457287, 5nmol; 4459405, 50 nmol; 4457289, 250 nmol)
Invitrogen In Vivo GAPDH Positive control siRNA (Cat. No. 4457288, 5 nmol; 4459407, 50 nmol, 4457291, 250 nmol)
Invitrogen In Vivo Factor VII Positive control siRNA (Cat. No. 4459408, 50 nmol; 4457292, 250 nmol)
Silencer Select Negative Control No. 1 siRNA, in vivo ready (Cat. No. 4404020, 250 nmol)
Silencer GAPDH Positive Control siRNA, in vivo ready (Cat. No. 4404025, 250 nmol)

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

How stable is Stealth siRNA, can I use it for in vivo research?

Stealth siRNA is markedly more stable in serum than unmodified siRNA. This increased stability is particularly beneficial in vivo where the RNAi may be exposed to more proteases. In collaboration with Intradigm, Stealth RNAi has been demonstrated to have activity in vivo when injected intra-tumorally.

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

How can I order in vivo siRNA duplexes?

Pre-designed Stealth RNAi and Ambion In Vivo siRNA can be ordered directly from our pre-designed siRNA search interface (https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai.html?open=faq#search) to target human, mouse, or rat genes.

Custom siRNA designs for in vivo experiments can be ordered from two interfaces, depending on the product format being requested: Custom Ambion In Vivo siRNA is ordered from the GeneAssist Custom siRNA Builder (https://www.thermofisher.com/order/custom-genomic-products/tools/sirna/), while Stealth RNAi is ordered using the BLOCK-iT RNAi Express for in vivo synthetics (https://rnaidesigner.thermofisher.com/rnaiexpress/inVivoService.jsp).

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

Do I need endotoxin testing for my in vivo RNAi duplexes?

It is recommended to reduce the chance of eliciting an immune response.

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

I want to order larger scales than the standard offering, whom do I contact?

Please contact rnairesearcher@invitrogen.com regarding large scale in vivo siRNA or fill out the quote request form (www.thermofisher.com/us/en/home/global/forms/custom-sirna-mirna-quote-request.html).

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

How should I deliver my in vivo RNAi molecules?

Several different approaches have been used for siRNA delivery, including various local delivery techniques and systemic delivery. We offer a lipid reagent, Invivofectamine 3.0 Transfection Reagent for in vivo siRNA systemic delivery. Learn more about this reagent here (https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai/invivofectamine-reagent.html).

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

Should I use siRNA or vectors for in vivo RNAi?

RNAi can be delivered using two different approaches-siRNA synthetic duplexes or siRNA expressed from plasmids or viral vectors (shRNA, miRNAi). siRNA are becoming the method of choice for the fast development of therapeutics. They are easy to use, easy to design, and easy to synthesize. siRNA can be rapidly identified and multiple genes can be targeted at the same time. With RNAi vectors, the expression will be steadier as a result of the possibility of stable integration of the plasmid into the genome, and they have the ability to target nondividing cells such as stem cells, lymphocytes and neurons. The drawbacks are the danger of oncogenic transformation from insertional mutagenesis, and unanticipated toxicity from long-term silencing of human genes and/or having high amounts of siRNA inside the cell (Grimm D. et al.: Nature 441: 537-541 (2006)).

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

I am planning on using siRNA for my in vivo experiments, what purity should they be?

For Stealth siRNA we recommend to select HPLC in vivo purity. For Invitrogen In Vivo siRNA, we recommend in vivo ready purity.

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

Should I use labeled or unlabeled siRNA for my in vivo siRNA experiments?

We have demonstrated that labeling Stealth and Silencer siRNA does not hamper their knockdown potency. An alternative approach is to mix unlabeled duplexes with labeled control duplexes; this method is more commonly used with in vivo siRNA, and allows progression to clinical research unhindered by questions about the possible effects of a label.

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

Should I use chemically modified duplexes for my in vivo siRNA experiments?

Chemically modified siRNA duplexes have a number of advantages over standard siRNA duplexes, including the minimization of off-target effects, enhanced stability, and reduced toxicity. For these reasons, chemically modified siRNA duplexes are recommended for in vivo siRNA experiments. For some pilot experiments, where the only aim is to determine biodistribution, unmodified siRNA and fluorescent controls are useful and slightly more cost-effective.

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

What information do I need to use the BLOCK-iT RNAi Designer?

Please provide either the accession number or nucleotide sequence. You will also have the choice of the region of the gene you would like to target: ORF, 5' UTR, or 3' UTR and the species. The percentage G/C content of the siRNA can also be chosen. Lastly, you can have your input sequence blasted against sequences in the database to ensure unique regions are used to design the siRNAs.

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

I would like to order a non-modified siRNA. What do you recommend?

You could order a Block-it RNA oligo or custom RNA oligo. Block-it RNA oligos are small 21-23 unmodified, nucleotide sequence-specific RNA duplexes. To design and order BLOCK-iT siRNAs, please visit the BLOCK-iT RNAi Designer (http://rnaidesigner.thermofisher.com/rnaiexpress/). Custom siRNAs can also be ordered by email (http://www.thermofisher.com/us/en/home/life-science/rnai/rnai-email-order-forms.html).

Custom RNA oligos can be ordered as single stranded oligos or duplexed to create a double stranded RNA. The oligo can be up to 50 bp in length, with the option of a biotin, fluorescein or phosphate 5' modification. These oligos will be delivered at desalted purity only. To order custom RNA oligos, visit this website (https://www.thermofisher.com/order/catalog/en/US/adirect/lt?cmd=ConfigureRNASingleTube).

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

I would like to order an in vivo purity or fluorescent labeled Stealth RNAi siRNA. Do you offer this as an option?

If in vivo purity and/or 5' sense label (Biotin, Alexa Fluor 488, 546, 555, 647, 680, 750) is required, you can order the siRNA through our RNAi in vivo Designer page (http://rnaidesigner.thermofisher.com/rnaiexpress/inVivoService.jsp). These siRNA can be ordered as HLPC or desalted in vivo purity or HPLC in vivo purity.

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