Gibson assembly interposition improves amplification efficiency of long DNA and multifragment overlap extension PCR.

For difficult overlap extension PCR, a Gibson assembly process was inserted between the two PCR rounds to facilitate the formation of complete gene templates at a moderate temperature. That is, after amplifying each DNA fragment, they were preluded by a Gibson assembly process in equal proportion. Then, the assembled mixture was used as a template for the second PCR round. This idea was tested and verified by taking the cloning example of a single and a double site mutation of the retinoblastoma gene. This scheme associates overlap extension PCR with Gibson assembly exquisitely, significantly improving gene amplification efficiency, particularly in the fusion of long genes and multifragments using overlap extension PCR.

All-fiber high-speed image detection enabled by deep learning.

Ultra-high-speed imaging serves as a foundation for modern science. While in biomedicine, optical-fiber-based endoscopy is often required for in vivo applications, the combination of high speed with the fiber endoscopy, which is vital for exploring transient biomedical phenomena, still confronts some challenges. We propose all-fiber imaging at high speeds, which is achieved based on the transformation of two-dimensional spatial information into one-dimensional temporal pulsed streams by leveraging high intermodal dispersion in a multimode fiber. Neural networks are trained to reconstruct images from the temporal waveforms. It can not only detect content-aware images with high quality, but also detect images of different kinds from the training images with slightly reduced quality. The fiber probe can detect micron-scale objects with a high frame rate (15.4 Mfps) and large frame depth (10,000). This scheme combines high speeds with high mechanical flexibility and integration and may stimulate future research exploring various phenomena in vivo.