Revolutionize Single-Cell Genome Analysis! AGM^™(Agarose Gel Microcapsules)

This kit is based on the Agarose Gel Microcapsules (AGM) single-cell containment technology^*, which utilizes an agarose gel shell. A microbial single-cell can be embeded in a fine AGM™ using standard biological lab equipment. This kit would significantly improve genome coverage rate which is a challenge in microbial single-cell whole genome analysis.
* Utilizes RIKEN patented technology (Patent No. 7018685)

Outline of AGM™(Agarose Gel Microcapsules)
Inside of AGM™ is liquid and the outside is covered with agarose. One AGM™ encapsulates one microbial cell, but no external contamination of cells or genomic DNA will occur. On the other hand, DNA polymerase, dNTPs and primers can pass through agarose shell, enabling single-cell DNA amplification within one capsule. Since AGM™ dissolves upon heating, the amplified DNA can be easily extracted. After extraction, it can be used for DNA library preparation and NGS analysis. Hundreds of thousands of AGM™ can be conveniently produced in a single experiment.

Conventional Method: after the cell was isolated and placed onto a PCR plate using flow cytometer, MDA^* reaction was applied.
AGM™: MDA^* reaction was applied to a single cell contained within AGM™.
＊MDA(Multiple Displacement Amplification): an isothermal DNA amplification method using Phi29 DNA polymerase and random primers in which complementary strand displacement reactions occur continuously at multiple sites.

Technologies for miniaturizing reaction volume to improve genome coverage, such as gel-bead-embedding (center figure) and emulsion (right figure/small droplets suspended in oil: Water-in-Oil), have been put to practical use, but there are problems including the restriction of spatial freedom due to the embedding of microbes in the gel, the inability to supply necessary components from outside, difficulty in isolating fragile emulsions, and the need for expensive and specialized equipment, labor-intensive work.
The AGM™, on the other hand, is a method that overcomes these problems.

First, prepare AGMs where cells from a microbial sample are individually embedded using the AGM™ Reagent Kit. Then, perform single-cell genome amplification in the AGM and prepare a DNA library. Conduct DNA sequencing of the library and analyze the microbial genomic DNA derived from single cell.

Fig. A : E. coli grown from single cell in AGM™ observed under phase-contrast microscope
Fig. B : E. coli DNA in Fig. A was stained with SYBR Green, and the fluorescent image and Fig. A were overlapped.

AGM™ containing E. coli single-cell was prepared. AGM™(20 μL) and LB medium(1 mL) in 15 ml tube were incubated at 37°C for 1 day. Growth of E. coli, which was single cell at the start of culture, was observed in the AGM™ (the above figure). It is expected that AGM™ can be used for the culture of single microbial cell.

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