Applied Biosystems  The 3730 XL Sequencer is a high-quality sequencing platform for long- segment read and sequence analysis with a wide range of applications . The 3730 XL can simultaneously analyze 96 samples. The instrument uses simultaneous detection of 4 colors of fluorescence, which can run 24 hours without interruption , automatic filling, loading, electrophoresis separation, detection and data analysis. In addition to the instrument to complete the sequencing of a new gene or comparative sequencing, various fragments may be analyzed, including analysis of microsatellite DNA, genotyping comparative study of single nucleotide polymorphism (SNP) and the like.
ABI 3730XL Sequencer
Sequencing peak map
Principle of dideoxy chain termination method
The dideoxy chain termination method, also known as the Sanger method, is based on the principle that DNA templates are replicated in the presence of DNA polymerase, primers, and four deoxynucleotide triphosphates (dNTPs) , respectively, in a four-tube reaction system. The ratio of four dideoxynucleoside triphosphates (ddNTPs) was introduced . Since ddNTP lacks the 3 ' -OH group required for extension , when ddNTP is incorporated into the end of the chain, the chain stops extending. Thus, a series of DNA fragments with ddNTPs at the 3 ' end are produced in each reaction system . After the reaction was terminated, gel electrophoresis was carried out in 4 lanes to separate DNA fragments of different lengths, and the adjacent fragments differed by one base in length. After autoradiography, the base sequence of the synthetic fragment can be obtained based on the dideoxynucleoside at the 3' end of the fragment.
S anger method sequencing schematic
ABI 3730XL sequencer technology principle
The ABI 3730XL Auto Sequencer is a DNA sequencer based on capillary electrophoresis and fluorescent labeling . Based on the Sanger method, the 3730XL sequencer has 96 capillaries and replaces isotopic labels with fluorescent labels, greatly increasing the speed and accuracy of DNA sequencing. The bases of the four dideoxynucleotides (ddNTPs) were labeled with different fluorescences . The four fluorophores on the DNA fragments of different lengths were excited by laser light when they passed through the capillary , emitting different colors of fluorescence and detected by CCD . The system recognizes and translates directly into DNA sequences.
Instrument characteristics
1. Double-beam double-sided laser excitation, grating spectroscopic device and post-ultra-thin CCD detection imaging system .
2. Automatic glue filling, sample loading, electrophoresis separation, detection and data analysis, continuous operation for 24 hours without manual intervention .
3. 96 samples were electrophoresed in one run, and 12 times of sequencing electrophoresis was performed in one day. The sequencing yield per day was 1152 samples .
4. Automatic base recognition, genotype analysis and quality score determination .
5. New liquid POP-7 separation gel, the capillary wall coating within the dynamic, the read sequence length of more than 1 0 00bp, accurate reading sequence of 800bp.
6. A good temperature control device provides a larger temperature control range ( 18~70 °C ) and an electrophoresis temperature of 70 °C , which helps to remove the influence of secondary structure .
7. High sensitivity reduces the amount of DNA and sequencing reagents used .
8. Higher automation reduces labor costs and costs of human error.
Technical application
1. Sequencing of bacterial solution, plasmid, and PCR product
2. Fosmid/Cosmid/BAC double-end sequencing
3. Large fragment shotgun sequencing
4. EST sequencing and analysis
5. Step sequencing
6. SNP discovery and resequencing
7. STR classification
Meiji Bio High-throughput Sequencing Technology Service Phone: 021-51872398-810
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