dCITI-Seq: droplet combinational indexed transposon insertion sequencing.

The rapid development of high-throughput parallel sequencing poses new challenges for large-scale barcoding and sequencing library construction. Here, we present droplet combinational indexed transposon insertion sequencing (dCITI-Seq), in which samples are indexed by the direct insertion of index-containing adaptors through transposition. The random combination of two sets of adaptors with known barcodes and massively parallel transposition was realized via a robust droplet pairing and merging platform. This strategy potentially enlarges the indexing capacity and decreases index crosstalk. Also, dCITI-Seq exhibited a lower GC base preference than conventional in-tube transposition library preparation. With a custom bioinformatic processing, it could be further applied to large-scale single-cell sequencing.

Quantifying genome DNA during whole-genome amplification via quantitative real-time multiple displacement amplification.

DNA quantification is important in the research of life sciences. In an independent quantification process, the extracted part of a DNA sample is usually difficult to be recycled for further use while the widely used real-time PCR is used to count the copies with certain sequences. Based on the popular multiple displacement amplification (MDA), we proposed and performed quantitative real-time MDA to obtain the information of template amount based on fluorescence signals while amplifying whole-genome DNA. The detection limit of real-time MDA was as low as 0.5 pg µl-1 (5 pg DNA input), offering the whole-genome research a promising tool to quantify the entire DNA during amplification without sacrificing sample completeness or introducing redundant steps.

Single-Molecular Förster Resonance Energy Transfer Measurement on Structures and Interactions of Biomolecules.

Single-molecule Förster resonance energy transfer (smFRET) inherits the strategy of measurement from the effective "spectroscopic ruler" FRET and can be utilized to observe molecular behaviors with relatively high throughput at nanometer scale. The simplicity in principle and configuration of smFRET make it easy to apply and couple with other technologies to comprehensively understand single-molecule dynamics in various application scenarios. Despite its widespread application, smFRET is continuously developing and novel studies based on the advanced platforms have been done. Here, we summarize some representative examples of smFRET research of recent years to exhibit the versatility and note typical strategies to further improve the performance of smFRET measurement on different biomolecules.