37 results about "Restriction Enzyme Recognition Site" patented technology

Provided are polynucleotides comprising a mobile bacterial island nucleotide sequence (B-INS) comprising one or more modifications (a modified B-INS) that include deletion or modification of restriction enzyme recognition sites to inhibit restriction enzyme cleavage of the B-INS, and an insertion of at least one cargo sequence into the B-INS. The B-INS is packaged within a bacterium into a phage-like particle that contains a bacteriophage capsid, tail and tail fiber proteins. The phage-like particle function as an antibacterial drone "ABD" and infects bacteria. The cargo sequence is introduced into bacteria infected by the ABD and kills the infected bacteria or inhibits growth of infected bacteria.

This invention discloses a method for constructing a next-generation sequencing library using long PCR amplicon sequences, comprising: performing PCR amplification on a target sequence using primers to obtain long PCR amplicon sequences; digesting and fragmenting the sequences using a mixed enzyme system, followed by end-repair and A addition, wherein the mixed enzyme system includes Exo I enzyme, UDG enzyme, and Taq DNA polymerase; ligating the A-added product to sequencing adapters to obtain adapter ligation products in the form of "adapter-DNA insert-adapter"; and performing a first round of purification to obtain a next-generation sequencing library, which can be used directly for sequencing or enriched by further PCR amplification before sequencing. This invention enables rapid construction of long PCR amplicon libraries for next-generation sequencing platforms, is simple to operate, and is suitable for large-scale applications. It uses UDG enzyme and dUTP to fragment the long PCR amplicon sequences, eliminating the need for specific restriction enzyme recognition sites and improving randomness.