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Longmers
of RNA up to a 80 mer can be efficiently produced by
RNA-TEC. These oligos are being used by our clients
for a variety of complex applications that require high
purity. Also shRNA, which is an efficient tool for RNA
interference, belongs to this category. For more info
on shRNA please click
here.
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Purity: All purities ranging from 90 to 98
% are available. The longmers
corresponds to 1 or 2 purifications. For every request
we design a custom made purification protocol to meet
the needs of the customer in a cost efficient way.
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Yield: The yield depends on the purity of the
longmers you require and the length of the longmers.
For some examples see below.
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Standard RNA & DNA: In our portfolio we
have the below mentioned scales of standard long RNA
and DNA oligos (longmers). RNA oligos up to lengths
of 45 bases are standard. For longer RNA oligos, longmers
please inquire.
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Typical products: All scales above 15 µmol
and purity levels up to 98 % are available on request.
Please note that for these specialty products RNA-TEC
will still provide the 1 micromole scale synthesis as
well.
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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 longmers 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 1 µmol scale synthesis of
a 58mer RNA oligo where a high purity was required resulted
in 1.2 mg or 65 nmole of pure product and a purity of
around 98 %. For chromatograms see below. This oligo
was first purified by reversed phase on which you can
first see a through peak (retention time approx. 3 min)
followed by the non-full length sequences (retention
time approx. 6 to 12 min) and finally the full length
trityl-on product (retention time 20 to 27 min). To
guarantee the high purity required an additional anion-exchange
HPLC purification was performed. The quality control
was performed on a high resolution analytical reversed
phase column.
A 14.8 µmol scale synthesis of a 40mer RNA oligo resulted
after 2 purifications in 10 mg or 0,79 µmol of pure
product. The purity as determined on analytical anion-exchange
was greater than 95 %.
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