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Automated Dideoxy Sequencing
CD Genomics' Sanger sequencing team provides researchers with automated dideoxy nucleotide-based dideoxy sequencing. We use ABI's 3730 DNA Analyzer for automated DNA sequencing based on capillary electrophoresis. In automated dideoxy sequencing, the dideoxynucleotides have been chemically modified to fluoresce under UV light (four different fluorescence colors for the four dideoxynucleotides A, G, C, and T), and the computer reads each band of the capillary gel sequentially, identifying each terminal ddNTP by its fluorescence. All users receive a color optical density scan showing the four different colored peaks corresponding to the different bases. Double-stranded DNA plasmids, BACs, single-stranded DNA, and PCR fragments are available for this sequencing service.
Customers Need to Provide
Double Stranded DNA
- 0.7-1.0 µg total
Single Stranded DNA
- 0.2-0.4 µg total
Cosmid, BAC
- 2.0-5.0 µg total
PCR Fragments
- 100-200 bp: 10 ng
- 200-500 bp: 10-20 ng
- 500-1000 bp: 20-50 ng
- 1000-2000 bp: 50-100 ng
Please note the purity of the samples, we require 260/280 = (1.8-2.0), 260/230 > 1. We reject samples containing salt, samples with ratios 260/280 out of range, samples containing RNA, samples with the slow growth of the host vector, and samples still containing ethanol or phenol.
Tubes and Plates
The total volume of the reaction should be:
- 15 µL when we add the primer.
- 20 µL when the sample already includes the primer. (If the DNA is concentrated, please dilute it in water up to the right volume)
- Please bring the sample in 0.6 mL tube.
- For more than 40 samples you can prepare them on 96 well PCR microplate (0.2 mL). In each well put 10 µL (total) contain half amount of the DNA and half amount of the primer.
- The samples should be from A1 - H1, A2 - H2, etc. (the rows are of 8 samples each).
We Are Ready to Provide
- After receiving your samples, we will mark your tubes or plates with a unique QR code.
- Then perform the appropriate sample quality inspection, processing, and purification based on your sample type.
- After complete sequence purification, automated Sanger sequencing was performed. After the sequencing is completed, we will conduct corresponding data analysis according to your research needs. Finally, we will return the obtained data and corresponding color densitometric scans to you.
CD Genomics, as a biotechnology company specializing in sequencing, provides you with a high-quality automated dideoxy sequencing service. If you need it, please contact us directly.
For research use only, not for any clinical use.
