Wednesday, March 4, 2015

Product Focus: RNA in vivo transfection reagent

work well for siRNA, miRNA, guide RNA and long ncRNAs

Description

Biotool RNA in vivo Transfection Reagent is a nanoscale polymer based reagent, successfully used to in vivo transfer siRNA, miRNA (mimic and inhibitor), guide RNA or long ncRNAs for research purposes such as gene functional studies or RNAi in vivo studies. This reagent interacts with nucleic acids and moves together into cells via endocytosis. In addition to efficiently transporting nucleic acids into the cells, the formula is also able to lead to the protection of the nucleic acids from degradation in vivo. Biotool RNA in vivo Transfection Reagent has been proven to be able to perform the maximum delivery of siRNA, miRNA (mimic and inhibitor), guide RNA or long ncRNAs into different organs in the body. No significant pro-inflammatory responses have been detected using this reagent.

Biotool RNA in vivo Transfection Reagent performs high transfection efficiency in vivo.


Biotool RNA in vivo Transfection Reagent test: Systemic administration (i.v.) of Biotool RNA in vivo Transfection Reagent conjugated with siRNA (siRNA/mouse: 1mg/kg, 2mg/kg and 3mg/kg) targeting coagulation factor VII (FVII) gene mRNA or non-silencing control siRNA (3mg/kg) following our protocol. Liver was collected and total RNA isolated 48 hours after tail vein injection. Samples were analyzed by qRT-PCR for FVII mRNA expression levels. β-Actin RNA levels were used to normalize the expression data. Data are means +/- SD (n=6).

No detectable inflammatory response.


Serum concentration of IL12/IL23 and IL6 following intravenous nucleic acid delivery using Biotool RNA in vivo Transfection Reagent or saline (negative control) 24h after injection respectively. The positive control is an IP injection of 50μg inactivated E.coli (LPS). 40ug siRNA were delivered with or without Biotool RNA in vivo Transfection Reagent.

Products
B45212     20 injections (0.5 ml)
B45215     100 injections (2.5 ml)
B45218     1000 injections (25 ml)


Flow Chart


 Troubleshooting


Problem Possible Reason Suggested Improvement
Precipitate formation in transfection mixture RNA not dissolved in distilled H2O (e.g. Tris-EDTA) Prepare fresh culture.
Excessive RNA concentration in transfection mixture Ensure RNA concentration in transfection mixture ≤ 0.5 µg/µL before injection
Transfection mixture froze or was stored at low temperature for a long period of time Freshly prepare transfection mixture and use immediately
Low transfection efficiency Low administration dose Scale up administration dose in the case of exclusion of animal lethality
Inflammation response or lethality of animal Injection manipulation error Inject the transfection mixture at correct site by correct method
Excessive RNA amount Scale down RNA amount and adjust the volume of transfection reagent accordingly
Endotoxin in transfection mixture Prepare endotoxin-free RNA
Pass transfection mixture through a 0.22 µM sterile filter before injection


 User Guide

Biotool RNA in vivo Transfection Reagent.pdf



Where can I find more information about Biotool?

Visit the manufacturer page at www.stratech.co.uk/biotool, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.
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