Creative Biogene can provide you with plasmid sequencing, amplicon sequencing, mRNA/cDNA sequencing, shotgun sequencing, Primer-Walking and viral DNA/RNA sequencing based on sanger sequencing methods. Our Sanger sequencing supports a wide range of DNA sequencing applications and we are dedicated to helping you find solutions in cancer research, genetic disease research, pathogen detection and analysis, epigenetic analysis, agrobiology research, food inspection and more. If you have any questions about submitting samples for Sanger sequencing or have any sequencing projects, please feel free to contact us.
Sanger sequencing, the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerases during in vitro DNA replication, is the most widely used method for detecting SNVs. Sanger sequencing is a "first-generation" DNA sequencing method. Despite the advantages of next-generation sequencing technologies with orders of magnitude higher throughput, Sanger sequencing still has an important place in clinical genomics.
Many clinical laboratories currently use the sanger method for direct sequencing of PCR products. In cycle sequencing, PCR products, sequencing primers (forward or reverse), deoxynucleotides (dNTPs), dideoxynucleotides (ddNTPs), and thermostable DNA polymerase are all included in the reaction mixture. First, the sequencing primers hybridize to the PCR product and are extended by DNA polymerase during the PCR process. ddNTPs are randomly incorporated into DNA strands during extension, terminating chain extension at every position in the sequence. Subsequent capillary electrophoresis separates the DNA strands by size and uses each fluorescent dye to identify the terminating nucleotide. Sanger sequencing is considered the gold standard method for mutation analysis, which can determine the entire sequence and identify unknown mutations.
Fig.1 The Sanger sequencing method in 7 steps.
At Creative Biogene, we accept a variety of templates to meet your Sanger sequencing needs. Over the past few years, our Sanger sequencing service has helped many researchers to elucidate a wide range of research questions, such as drug resistance detection and tumor mutations when SNPs are associated with specific genomic regions. Our simple and fast Sanger sequencing turnkey solution provides the sample-to-result process. A complete list of workflows and acceptable sample types is listed below.
Prepare high-quality sequencing templates and primers for setting up sequencing reactions. PCR templates require purification or cleanup to remove dNTPS, primers, and DNA polymerase.
Amplifies linearly using one primer and one template and generates nested fluorescently labeled extension products with single nucleotide differences.
Cleanup of completed sequencing reactions to remove unincorporated dyes and other reaction components from end-labeled extension products.
Loaded or unresuspended extension products on a CE instrument to detect fluorescent signals, which are converted to DNA sequences using base calling software.
As a complementary workflow step for instrument maintenance, after hundreds of CE runs, the capillary array is regenerated to remove contaminants accumulated on the array. The regenerated array regains optimal performance.
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