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Integrated DNA
Technologies, BVBA
Provisorium 2
Minderbroedersstraat 17-19
B-3000 Leuven
BELGIUM
Tel: +32-16-337096
Fax: +32-16-337097
info@rna-tec.com
How to reach us
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siRNA has rapidly become a widely
used tool for determining gene function and validating potential
therapeutic targets both in cell culture and in vivo.
With the present rate of progress and most recent animal experiments
siRNA
is poised to become a new generation of rationally designed
drugs. However, this will need sophisticated delivery techniques
for the siRNA to reach and enter the correct
cells and tissues. For some key references see below.
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Purity: We only supply fully purified and desalted
RNA even though we have very good crude synthesis based on
HPLC (see chromatogram below) ready to use as siRNA . The
reason for this is to avoid misleading results in biological
experiments that could be caused by traces of contaminants
present in non-purified samples. Our standard purity is greater
than 95%, but on average about 98 %.
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Yield: RNA-TEC performs a 15 to 20 micromole scale
synthesis to guarantee 25 mg of pure product from each strand.
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Typical products: For in vivo siRNA experiments
for target validation we are providing as standards 25 mg
or 50 mg of each strand for a very competitive price. Other
scales are available on request.
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Transcribed RNA versus chemically synthesized RNA:
If you are planning to do in vivo experiments in which
you will need more of the siRNA
please bear in mind that chemical synthesis will be much
more cost effective. This is largely due to the cost of the
T7 RNA polymerase. Moreover, some sequences are difficult
to transcribe due to their secondary structure and sequence
whereas chemical synthesis does not suffer from such limitations.
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Caplen, N. J., Parrish, S., Imani, F., Fire,
A. and Morgan R.A. (2001). Specific inhibition of gene
expression by small double-stranded RNAs in invertebrate and
vertebrate systems. PNAS 98, 9742-9747. [Free
full text]
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Harborth, J., Elbashir, S.M., Bechert, K.,
Tuschl, T. and Weber, K. (2001). Identification of essential
genes in cultured mammalian cells using small interfering
RNAs. J Cell Sci 114 , 4557-4565. [Free
full text]
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McCaffrey, A. P., Meuse, L., Pham, TT. T.,
Conklin, D.S., Hannon, G. J. and Kay, M. A. (2002). RNA
interference in adult mice. Nature 418, 38-39.
[Abstract]
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Lewis, DL., Hagstrom, JE., Loomis, AG., Wolff,
JA., and Herweijer, H. (2002). Efficient delivery of
siRNA for inhibition of gene expression in postnatal mice.
Nat Genet 32, 107-108. [Abstract]
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Hasuwa, H., Kaseda, K., Einarsdottir, T.
and Okabe, M. (2002). Small interfering RNA and gene
silencing in transgenic mice and rats. FEBS Lett 532,
227-230. [Abstract]
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Couzin J. (2003). Mini RNA molecules
shield mouse liver from hepatitis. Science 299,
995.
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Song, E., Lee, SK., Wang, J., Ince, N., Ouyang,
N., Min, J., Chen, J., Shankar, P. and Lieberman, J.
(2003). RNA interference targeting Fas protects mice from
fulminant hepatitis. Nat Med Feb 10 [Abstract]
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Hemann, MT., Fridman, JS., Zilfou, JT., Hernando,
E., Paddison, PJ., Cordon-Cardo, C., Hannon, GJ. and Lowe,
SW. (2003). An epi-allelic series of p53 hypomorphs created
by stable RNAi produces distinct tumor phenotypes in vivo.
Nat Genet Feb 3 [Abstract]
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Yield examples: An 18.6 µmol scale synthesis of a
21 mer RNA resulted in 31 mg or 5 µmol of pure product after
a single anion-exchange HPLC purification followed by a desalting.
The purity was > 98% as determined by analytical anion-exchange
HPLC (see below).
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Important notice:
Purchasers of RNA products intended for use in RNA interference
experiments, whether in vitro or in vivo may require a license
to one or more of the granted patents and pending patent applications
covering the usage of siRNA for gene knockout, target validation
and therapeutic development. Particularly relevant are patents
granted to Ribopharma AG (EP 1144623 A1) and the Carnegie
Institute of Washington (US 6,506,559 B1) and applications
(including WO 01/75164 A2) filed by the Whitehead Institute
of Biomedical Research, the Max-Planck-Gesellschaft zur Forderung
der Wissenschaften e.V., the Massachusetts Institute of Technology
and the University of Massachusetts Medical Center. If in
doubt please contact the owners of the patents directly.
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shRNA (short hairpin RNAs) are
able to silence genes as efficiently as their siRNA counterparts . Moreover, shRNAs exhibits better reassociation
kinetics in vivo than equivalent duplexes.
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Purity: We only supply fully purified and desalted
RNA even though we have very good crude synthesis based on
HPLC (see chromatogram below). The reason for this is to avoid
misleading results in biological experiments that could be
caused by traces of contaminants present in non-purified samples.
Our standard purity is around 95%. Due to the length of these
oligos (between 50 and 60 mer) RNA-TEC utilizes special HPLC
purification methods which made it possible to reach this
purity in a single step, which means that losses of product
resulting from multiple purification steps can be avoided.
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Yield: RNA-TEC performs a 15 to 20 micromol scale
to guarantee 10 mg of pure product.
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Typical products: For in vivo shRNA experiments
for target validation we are providing as standard 10 mg or
25 mg of pure product for a very competitive price. Other
scales are available on request.
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Transcribed RNA versus chemically synthesized RNA:
If you are planning to do in vivo experiments where
you will need more of the shRNA please bear in mind that chemical
synthesis will be much more cost effective. This is largely
due to the cost of the T7 RNA polymerase. Moreover, some sequences
are difficult to transcribe due to their secondary structure
and sequence whereas chemical synthesis does not suffer from
such limitations.
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Yield examples: A 14.9 µmol scale synthesis
of a 56 mer RNA resulted in 19.5 mg or 1.1 µmol of pure
product after purification and desalting. The purity was >
95% as determined by analytical anion-exchange HPLC. To reach
this purity and yield RNA-TEC follows a completely customized
purification protocol for these long RNA oligos. The purity
> 95 % (see analytical anion-exchange HPLC) is reached
after a single purification, is this case an anion-exchange
purification, as shown below. This product was produced as
a guaranteed 10 mg of shRNA oligo.
RNA-TEC: siRNA .
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