cDNA Library Construction
NorClone offers custom cDNA library construction service.
We have
extensive experience in constructing high quality cDNA libraries in a
vector of choice. The cDNA library construction can begin with the
following customer provided starting materials:
Starting Material
|
Amount
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Tissue
|
≥ 1 gram
|
Cells
|
1 x 108
|
Total RNA
|
≥ 1 mg
|
mRNA
|
≥ 5 µg
|
The cDNA library is produced by using full-length enriched
and high quality size-fractionated cDNA. High-quality mRNA is synthesized
into cDNA using oligo-dT primers and the double stranded cDNA is
directionally cloned into a vector of choice. QC analysis is conducted to
confirm size and/or sequence identity of randomly sampled clones, as well
as overall cloning efficiency.
Shotgun
Library Construction & Sequencing
NorClone offers highly
flexible genomic shotgun library construction services, including shotgun
sequencing of the completed library. Shotgun library construction and
sequencing can begin with a variety of samples such as cosmid, BAC, and
YAC DNA.
Construction:
-
Plasmid DNA purification and/or quality analysis of
in-coming DNA
-
Random shearing to produce a population of DNA fragments
(typically 1-3 KB)
-
Ligation into a standard cloning vector and transformation
-
Libraries submitted to customer as plates, glycerol stocks,
or purified DNA
-
QC by PCR or DNA sequencing
Sequencing:
-
Plating, picking, and arranging colonies into 96- or
384-well plate format
-
Plasmid purification of clones
-
High-throughput sequencing
-
Contig assembly, mapping and gap closure
The project report is provided with the original sequencing
chromatograms, all primer sequence information, and the full consensus
sequence. All plasmid DNA and primers are delivered.
Please contact us for a
detailed quote or for more information.
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