The invention relates to the technical field of molecular biology and in particular relates to a novel DNA library construction kit and an application thereof. The DNA library construction kit provided by the invention comprises an adhesive end joint, a ligase, a DNA extension reagent and a DNA amplification reagent. The DNA library construction kit disclosed by the invention can be applied to a sample with low initial mass and can directly mediate the connection of DNA and the joint by a common ligase; and compared with previous methods, the kit has the advantages that the connection efficiency of samples is greatly increased, the diversity of the samples are retained to the greater extent, the stability is good, and library bias caused by artificial factors is shortened.

Novel DNA library construction method and application thereof

The invention discloses a single-cell methylation sequencing technology and an application thereof. The technology has the following advantages: (1) the technology has the characteristics of simplicity in operation, short manual operating time, and no pollution; (2) a pre-amplification primer is designed for the characteristics of a bisulfate treated genome sequence, the 3' end of the pre-amplification primer is an H base, and the H base does not complement or bind to C, so binding of the pre-amplification primer to the bisulfite converted sequence is not affected, and the probability of binding of a random primer and a linker sequence is reduced; and (3) the technology has a high Mapping rate, and effectively improves the use rate of sequencing data, and the use rate is 20-30% higher thanthe use rate (average 25%) of existing scRRBS and the use rate (average 10-30%) of scBS. The technology is of great significance for the study of single-cell DNA methylation.

Single-cell methylation sequencing technology and application thereof

The invention provides a kit suitable for whole-genome amplification of single cells. The kit can be effectively applied to a fixed single cell sample through primer optimization and amplification steps; and enzyme treatment for specific removal of uridylate can be used to the amplification of the single cell sample, so the error rate of amplification is significantly reduced. Compared with kits in the prior art, the kit in the invention has the advantages of high efficiency, simplicity, practicality, accurateness, small loss, low cost, good method repeatability, low error rate, high suitableness for the single cell whole-genome amplification kit of fixed samples, expansion of the application range of the samples, reduction of the error rate, and improvement of the detection precision.

Kit for whole-genome amplification of single cells

The invention relates to a construction method and application of a single-cell methylation sequencing library. The invention applies a multiple displacement amplification technology to construction of the single-cell methylation sequencing library, and richness and comparison rate of the sequencing library are obviously enhanced. The construction method comprises the following specific steps: firstly producing small segments containing uracil by utilizing an optimized bisulfate conversion kit, then carrying out amplification extension for multiple times by using a label-free random primer, degrading a free single-stranded small segment primer, then denaturalizing a paired double strand by utilizing high temperature, then connecting single-stranded small segments into a single-stranded long fragment, carrying out multiple displacement amplification by utilizing a label-free random primer segment with a terminal 5' closed, obtaining an amplification end product, and finally carrying outlibrary construction based on Tn5 enzyme on a reaction product.

Construction method and application of single-cell methylation sequencing library

The invention relates to the technical field of molecular biology and particularly relates to a novel methylation library-establishing kit and an application thereof. The novel methylation library-establishing kit at least comprises a bisulfate conversion reagent, a sticky end joint, ligase, a DNA extending reagent and a DNA amplifying reagent. The novel methylation library-establishing kit can beapplied to a sample with a low initial mass (as low as 6pg) and suitable for genome DNA with relatively poor mass extracted from FFPE samples and the like, so that the loss of sample information caused by DNA breakage during drug treatment is greatly reduced; and a double-chain DNA joint with a sticky end can be directly introduced by virtue of relatively economic and practical common ligase, sothat the accuracy of data can be improved, the diversity of samples can be preserved to a great extent, and the library deviation property caused by human factors can be reduced.

Novel methylation library-establishing kit and application thereof

The invention relates to the technical field of molecular biology and in particular discloses a library establishment method applicable to single cell genome methylation sequencing and application thereof. The library establishment method applicable to the single cell genome methylation sequencing, provided by the invention, comprises the following steps: (1) carrying out bisulfite transformation on a genome DNA (Deoxyribonucleic Acid) of a sample; (2) carrying out linear amplification on the transformed genome DNA in step (1); (3) carrying out exponential amplification on an amplicon obtained by the linear amplification in step (2), wherein the amplicon subjected to the exponential amplification is used as a sequencing library. According to the library establishment method disclosed by the invention, the starting amount of the genome DNA of the sample can be reduced to a pg grade; the library established by adopting the method is used for sequencing and most of cytosine of a whole genome can be detected; the library can cover most of regions of the whole genome.

Library establishment method applicable to single cell genome methylation sequencing and application thereof

The invention relates to the technical field of molecular biology and in particular relates to a novel methylation library construction method and an application thereof. The novel methylation libraryconstruction method provided by the invention is suitable for single-strand connection, amplification or library construction of a DNA sample treated by using bisulfite. Due to the adoption of a method for bisulfite treatment and then adhesive end joint connection, sample information loss caused by DNA breakage resulted from drug treatment is greatly reduced.

Novel methylation library construction method and application thereof

The invention provides an optimized single-cell transcriptome library establishment method. The method comprises the following steps: (1) obtaining a sample RNA through cell lysis or extraction; (2) performing reverse transcription operation on the sample to make the RNA form cDNA through reverse transcription; and (3) constructing a transcriptome library by using a separated cDNA, wherein the step (3) of constructing the transcriptome library by using the separated cDNA comprises the following steps: (3-1) performing a tail adding reaction or adopting a single-chain connection manner to add an adaptor; (3-2) performing two-chain synthesis on the cDNA and performing an amplification reaction; (3-3) performing exponential amplification; and (3-4) after transcriptome library construction isperformed on the cDNA amplification product subjected to step (3-3) treatment, removing ''useless DNA''; or after the ''useless DNA'' in the cDNA amplification product subjected to step (3-3) treatment, constructing a transcriptome library. Compared with the prior art, the method provided by the invention is more high-efficiency, simpler, more practical and more accurate, has less pollution, lessloss, low costs and good method repeatability, expands the application range of the sample and improves the precision degree of detection.

Single-cell transcriptome library establishment method and application thereof

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Library establishment method applicable to single cell genome methylation sequencing and application thereof

Novel methylation library construction method and application thereof

Single-cell transcriptome library establishment method and application thereof