Automated DNA Library Prep | 16S Amplicon Sequencing-Fecal DNA Samples

16S amplicon library construction involves extracting total DNA from a sample, performing targeted amplification of the hypervariable regions of the 16S rRNA gene using specific primers, and introducing sequencing adapters and sample-specific barcodes to generate a standardized sequencing library. This culture-independent approach enables efficient, high-throughput profiling of bacterial and archaeal community structures in complex environments.

Applications of Soil 16S Amplicon Sequencing

Reagent List for the Experiment

Category	Cat.No.	Product name
16S Amplicon Library Preparation	12933ES	16S (V4) Bacterial AmpSeq Prep Kit For MGI
Magnetic Beads	12601ES	Hieff NGS™ DNA selection Beads (Superior Ampure XP alternative)
Quantification	12642ES	1× dsDNA HS Assay Kit dsDNA qubit
Adapters	12330ES	Hieff NGS™ Unique Dual Barcode Primer Kit for MGI
User-Supplied Materials	—	Absolute Ethanol

The Automated Workflow

Figure 1. End-to-end automated 16S (V4) amplicon library construction workflow for soil DNA on the MGISP-960.

Pre-Experiment Preparation

1. Equilibrate magnetic beads to room temperature before use.

2. Prepare 80% ethanol.

3. Prepare 2 herbal medicine samples and 1 rapeseed sample, each containing 400 ng of DNA. The extracted DNA shows visible pigment contamination.

Protocol and Results

Parameter	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5
Sample Type	Soil	Soil	Soil	Soil	Soil
Input Amount	10 ng	10 ng	10 ng	10 ng	10 ng
Nucleic Acid Concentration (ng/μL)	33.4	107	92.2	79.6	161
PCR Master Mix	12.5 μL	12.5 μL	12.5 μL	12.5 μL	12.5 μL
16S Primer Mix	2 μL	2 μL	2 μL	2 μL	2 μL
Amplification Cycles	28
1st Purification	22.5 μL beads; 25 μL nuclease-free water elution
2nd PCR Input	3 μL of 1st purification product
Hieff Canace Amplification Mix	12.5 μL	12.5 μL	12.5 μL	12.5 μL	12.5 μL
Primer	2 μL	2 μL	2 μL	2 μL	2 μL
Amplification Cycles	6
2nd Purification	22.5 μL beads; 25 μL nuclease-free water elution
Library Concentration (ng/μL)	90.8	102	104	97.4	92.4

Agarose gel electrophoresis image of the libraries

The workflow is fully supported by a coordinated Yeasen product range, covering every step from DNA extraction through indexed library QC. All reagents and consumables are pre-validated for compatibility with the MGISP-960 deck layout described in this protocol.

Yeasen Product Portfolio for 16S Amplicom sequencing