High
Quality 454-Ready cDNA is made from our custom or catalog cDNA libraries.
Briefly, library DNA is digested with Not I and in
vitro RNA transcripts are produced using the SP6 RNA polymerase
promoter. Then, first strand cDNA is made from these transcripts
using a modified primer adapter that reduces the size of the poly
A sequence (to about 20 As). After the second strand is synthesized,
the double stranded (ds) cDNA is blunt ended and size fractionated.
This ds cDNA is now ready to be used with the 454 sequencing method
or with other next generation sequencing technology (i.e., illumina).
We guarantee at least 6 µg of ds cDNA, 150 ng/µl concentration
in TE and 1 kb average cDNA size, however, we generally produce
8 µg of cDNA, 200 ng/µl concentration and 1.5 kb average
cDNA size.
We will make 454-ready cDNA from
our custom or catalog libraries or directly from customer supplied
cDNA libraries, tissue or RNA.
Features
list:
-
High quality size fractionated cDNA.
- Reduced poly A size to improve sequencing results.
- Large amount of cDNA can be produced from nanoquantity levels
of tissue or RNA.
- Ideal subtrate for next generation sequencing methods.
Quality
testing: Gel analysis of cDNA population.
Shipment
and storage conditions: 454-ready cDNA will be shipped
in dry ice. Storage of cDNA at -80°C is recommended.
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