Description

Sanger Sequencing of Purified PCR Product/Plasmid Dextrose Technologies Pvt. Ltd. provides Sanger sequencing of purified PCR products and plasmid DNA for sequence verification, target confirmation, and molecular biology research. The service generates nucleotide sequence data accompanied by chromatogram files, allowing researchers to examine base calls and assess sequence quality. Whether you need to verify an amplified DNA fragment or confirm a selected region within a plasmid, Sanger sequencing provides a practical approach to targeted DNA sequence analysis. Applications • Verification of purified PCR products against an expected DNA sequence. • Confirmation of cloned inserts and selected plasmid regions. • Sequence confirmation following molecular cloning experiments. • Examination of nucleotide changes within the sequenced region. • Generation of sequence data for downstream alignment, identification, and comparative research. Service Overview Purified PCR product or plasmid DNA is submitted with an appropriate sequencing primer. Sequencing produces a chromatogram showing fluorescent signals associated with nucleotide base calls, together with a readable DNA sequence. Each sequencing reaction reads from the supplied primer in one direction. For confirmation from both directions, forward and reverse sequencing requirements should be discussed during submission. A single Sanger read does not cover an entire plasmid unless the region of interest falls within the obtainable read length. Sample Requirements • Sample type: Purified PCR product or plasmid DNA. • Sample volume: 10–20 µL per sample. • DNA concentration: 50–100 ng/µL. • Sequencing primer concentration: 10 pmol/µL. • Sequencing primer volume: 10 µL. Submission Information Clearly label each DNA sample and its corresponding primer. Provide the sample identifiers, sample type, and primer-to-sample mapping to help ensure correct processing. Include the expected PCR product size or plasmid details, where available. Submit purified DNA suitable for sequencing. Mixed PCR products, residual contaminants, or unsuitable primers can affect chromatogram quality and the usable sequence length. Discuss samples with uncertain purity or concentration before submission. Deliverables • Q20 read length of up to 800 bases, subject to sample quality and sequence characteristics. • AB1 chromatogram file for reviewing sequencing traces and base calls. • FASTA file containing the nucleotide sequence for downstream analysis. • Sequencing output in PDF format. Q20 indicates an estimated base-call accuracy of 99%. The usable high-quality read length may vary with template purity, primer performance, and sequence complexity. The stated maximum read length is not guaranteed for every sample. Turnaround Time 2–3 days, as listed in our service catalogue. Service Coverage Based in Bengaluru, Dextrose Technologies supports academic researchers, biotechnology companies, and industrial R&D teams across India, including Kochi, Kolkata, Delhi, Mumbai, Hyderabad, and Chennai. Contact Us Share your sample details and sequencing requirements for submission guidance. Dextrose Technologies Pvt. Ltd. Phone: +91 9880405558 Email: info.dextrosetech@gmail.com Website: www.dextrosetech.com