I. Gene Cloning
Subcloning
Gene Mutagenesis
siRNA and miRNA
CRISPR Cas9
Knockout and Knockin Vector Construction
II. Gene Discovery
Library Construction
Library Screening
5' and 3' RACE
Gene Synthesis
Gene Isoforms
III. Protein-Protein Interaction
Yeast 2 Hybrid Screen
Protein Expression
IV. Cell Culture and Recombinant Virus
Stable Cell Line
Virus Production
V. Molecular Diagnostics
Mouse Genotyping
Bacteria and Fungi Identification
 




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

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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