Direct Amplicon Analysis
Direct sequencing of purified PCR products to analyze amplicon diversity.
Direct, deep sequencing of purified PCR amplicon libraries (from 100 to 550 bp), using state-of-the-art Illumina® NGS platform, with bioinformatics analysis included.
Service Description
- Construction of an NGS library from PCR amplicon sample with all required QC steps
- Sequencing of the library with Illumina short reads, generating comprehensive coverage with high depth and accuracy
- Demultiplexing, trimming, and quality-filtering of raw sequencing reads
- Reference-based sequence binning of reads to analyze target abundance and frequency, or de novo population diversity analysis (including OUT analysis of metagenomic 16S/ITS libraries)
Bioinformatics Analysis
Standard types of bioinformatics analysis are included with NGS services.
Diversity Analysis
Reads are binned and aligned to target sequences to detect abundance and frequency of the target amplicons, or quantify diversity of amplicon population without target sequences.
Metagenomics Analysis
Reads from 16S and/or ITS target PCR samples are aligned to known species in public 16S/ITS databases to cluster for Operational Taxonomic Units (OTUs), detect population frequency, and identify taxonomy of microorganisms.
Custom Analysis
Complex bioinformatics analysis for various research purposes can be arranged as a custom project.
Acceptable Target Types
PCR Products (100bp to 550 bp in length)
Purified DNA Requirements
Please provide at least 10 µL, with OD260/280 >1.6 and <2.2, and all samples normalized to 3 ng/µL.
ACGT recommends DNA quantification using fluorometry. Quantification using Nanodrop is discouraged. Data quality of samples failing specified quantity and quality requirements cannot be guaranteed.
Bacterial Colony
Please provide sample colonies on agar plates, with 1 plate/sample.
Bacterial Culture
Please provide sample culture in tubes or plates, with at least 0.2 mL per sample.
For bacterial colony and bacterial culture samples, additional $50 per sample applies for plasmid template amplification.
Highlights
- High quality NGS reads at Q30 or higher (>99.9% accuracy)
- > 100X average coverage of target sequences
- 2-week turnaround time in most cases
- Detect amplicon sequence deviations at nucleotide-level sensitivity
- Most practical solution for amplicon population diversity analysis
- Standard bioinformatics analysis included at no additional cost
- 24/7 online ordering and data retrieval
- Dedicated technical support staff
- Free overnight shipping via ACGT drop boxes
- Real-time project tracking
Submitting Your Samples
Refer to DNA requirements for submission guidelines.
