53 results about "Transcriptome profiling" patented technology

This application discloses methods for cDN'A synthesis with improved reverse transcription, template switching and preamplification to increase both yield and average length of cDNA libraries generated from individual cells. The new methods include exchanging a single nucleoside residue for a locked nucleic acid (INA) at the TSO 3' end, using a methyl group donor, and / or a MgCb concentration higher than conventionally used. Single-cell transcriptome analyses incorporating these differences have full-length coverage, improved sensitivity and accuracy, have less bias and are more amendable to cost-effective automation. The invention also provides cDNA molecules comprising a locked nucleic acid at the 3'-end, compositions and cDNA libraries comprising these cDNA molecules, and methods for single-cell transcriptome profiling.

The invention belongs to the field of transcriptome analysis, and relates to a method for rapidly performing single-cell RNA reverse transcription and library construction. The invention can start from 1-2000 cells or 10pg-20ng of extracted eukaryotic total RNA within 5-6 hours, amplify 20-500ng high-quality full-length double-stranded cDNA, and obtain downstream High-quality cDNA libraries required for analysis. This invention effectively avoids 3' bias and genomic DNA contamination during cDNA synthesis. Expression quantitative molecular tags can assist in the calculation of gene expression. At the same time, the expression quantitative molecular tags can also retain strands while completely amplifying RNA sequence information. source information. The invention can obtain more than 95% success rate of reverse transcription and amplification library construction, the cDNA library can be seamlessly connected to the mainstream sequencing platform of Illumina, and the off-machine data (5M Reads) can detect more than 90% gene expression, and the gene expression consistency More than 90%, the amplification has no obvious bias, and the required sample input is less.

The invention discloses an application of m6A modification-related gene ALKBH5 in the preparation of medicines for treating diseases related to nerve axon damage. The invention also discloses an experimental verification of the application of m6A modification-related gene ALKBH5 in promoting the repair of nerve axon damage, which is characterized in that it comprises the following steps: S1, detecting the expression and location of ALKBH5; S2, interfering with the expression of ALKBH5 on nerve The effect of the axon growth; S3, the effect of interfering with the expression of ALKBH5 on the axon growth of the sciatic nerve; S4, the process of screening and functional testing of the ALKBH5 target gene LPIN2. The present invention verifies that interfering with ALKBH5 gene both in vivo and in vitro can significantly promote the growth of neuron axon, and promote the recovery of sciatic nerve function; and through transcriptome analysis and target gene verification after ALKBH5 interference, the target gene LPIN2 regulated by ALKBH5 is found and verified in Role in regulation of neuronal axon outgrowth.

Belonging to the genetic engineering and fermentation engineering field, the invention provides a method for increasing streptomyces secondary metabolite yield. The method includes the steps of: utilizing an induction promoter to control the expression of a target secondary metabolite biosynthetic gene cluster, and determining the optimal induction condition by means of a response surface model; conducting transcriptome analysis to screen a physiological promoter with consistent control behavior to the induction promoter; and finally, preparing plasmid utilizing the physiological promoter to control the expression of the target secondary metabolite biosynthetic gene cluster, and transferring the plasmid into host bacteria for fermentation. The method provided by the invention can get rid of dependency on an inducer, and realizes self-regulation of target biosynthetic gene cluster expression and increase of the target product yield. The method provided by the invention regulates the expression of secondary metabolite biosynthetic gene cluster from the dimensions of time and intensity, makes the expression fit the physiological metabolic behaviors of the host, and is very important for increasing the yield of the target secondary metabolite in streptomyces.