3 results about "Small fragment" patented technology

The present invention discloses a method for enriching small fragment cDNA of single cell sequencing for identifying miRNA expression in single cell transcriptome, using SPRISlect magnetic beads to sort library fragments, adding PEG10000, isopropanol and trichlorohexammine cobalt complex to the magnetic bead binding solution to improve the enrichment of small fragments of 100 to 300 bp. The present invention innovatively integrates DNA linear amplification and magnetic bead small fragment sorting technology with 10xGenomics single cell library construction method, solving the problem of small fragment PrimRNA data loss caused by insufficient small fragment enrichment ability in the single cell sequencing process in the prior art. The present invention has broad application prospects.

The invention relates to the technical field of gene engineering, in particular to a plasmid point mutation or small fragment insertion method, which is characterized in that a specific primer is designed at a mutation or insertion site, another primer is designed in other non-target regions of a plasmid to form upstream and downstream pairing with the specific primer, and the distance between the other non-target regions and the insertion site is 500-1000bp; carrying out a first-step PCR reaction on the plasmid by using the specific primer and the other primer; a PCR reaction product is recovered, and a DNA double-chain fragment is obtained; and carrying out a second-step PCR reaction on the plasmid by taking a DNA double-chain fragment as a large primer, and recovering and digesting a PCR product, so as to convert. The method can realize 100% correct rate of plasmid point mutation or small fragment insertion, is an efficient method for plasmid point mutation or small fragment insertion, and has a good promotion effect on molecular biology.