Uncloned Normalized cDNA (for 454 sequencing)

$3,299 US for 5ug of amplified uncloned normalized cDNA

 

Bio S&T delivers the highest quality cDNA for 454 sequencing (FLX and Titanium) and other downstream applications.

Our uncloned normalized cDNA is created using proprietary technology which features full-length-enriched and high quality size-fractionated cDNA (see Figure 1). We use a modified SMART method. We provide 5ug as a standard but higher amounts may be ordered at a rate of $100 per additional ug. A discount is available for large orders.

 

NEW ! NGS using Illumina Hi-seq 2000 platform and downstream bioinformatics analysis are available. Please contact us for details.

 

Type of library	Starting material	PrPrimers used
Full-length (eukaryotic)	Eukaryotic total RNA	Oligo dT  (+ interspersed dG to limit homopolymer regions)
Exclusively 5’ end	Eukaryotic mRNA	Random primers
Exclusively 3’ end	Eukaryotic mRNA	Oligo dT  (+ interspersed dG to limit homopolymer regions if desired)
Full-length (prokaryotic)	Prokaryotic total RNA	Random primers

Figure 1: Gel picture of a recently constructed plant normalized cDNA library (a portion of the uncloned cDNA was cloned).

 

1. 1ug of amplified non-normalized cDNA (available upon request).
2. 5ug of amplified uncloned normalized cDNA (suitable for directional cloning).
3. Primers for amplification (available upon request).

1. Option #1: Cloned cDNA library -Additional $250 US
   1. Bio S&T will test 10 clones following normalization for insert size determination (gel picture will be provided).
   2. 50,000 clones will be created (cDNA inserted into vector) and shipped as a glycerol stock.
   3. Additional clones can be provided for an additional fee. Please mention required number of clones when inquiring.
2. Option #2: Pooling - Additional $125 US (with purchase of Option #1)
   1. Cloned cells will be distributed in 1 x 96-well plate (about 500 clones/well).
3. Option #3: Screening - Additional $500 US (with purchase of Option #1)
   1. Screening of 50,000 clones using client-provided PCR primer pair. One positive cDNA clone will be provided.
4. Option #4: Sequencing sampler - Additional $200 US
   1. Sequencing (1 single pass/clone) will be performed 10 randomly selected clones. Data (chromatogram and text file) will be provided.
5. Option #5: + ug of uncloned cDNA - Additional $100 US/ug
   1. Purchase additional quantity of uncloned cDNA for $100 per microgram.

At least 1ug of total RNA (more is recommended if material is not limiting). RNA may be isolated using any standard method (TRIZOL protocol is highly recommended). Should material be limiting, please email us to obtain our RNA isolation protocol (provided to client once order is placed). Tissue or cells may also be provided (RNA isolation fee of $300 US will apply).

 

Please precipitate the total RNA and wash with 80% EtOH. Re-suspend pellet in 80% EtOH. It is important to:

1. Use a tube with a screw-cap microtube.
2. Fill the tube almost to the top with 80% EtOH.
3. Place parafilm around cap to prevent evaporation.

We recommend dry ice shipping but should this not be an available option, please send the material at room temperature via the courier of your choice to:

                    Bio S&T Inc.
                    Attn: Areti Karadimos
                    5020 Fairway Street, Suite 220
                    Montreal (Quebec) H8T 1B8, Canada
                    Tel: (514) 633-6006

We can also accept dry ice shipments. We have extensive worldwide shipping experience and can accept samples from any worldwide location.

 

Many of our clients have opted to use EnGenCore, a core genomic sequencing facility using the Roche 454 platform. EnGenCore is located at the Public Health Research Center at the University of South Carolina. To obtain a quote please contact engencore.sc.edu (1-803-777-4338)