The invention relates to the technical field of biotechnology and DNA detection, in particular to a high-sensitivity double-chain circulating tumor DNA detection method and a kit. The method disclosedby the invention comprises the following steps: carrying out DNA tail end repairing and tail end tailing reaction on a plasma free DNA, carrying out a connection reaction with a linker containing UMI, carrying out AMPpure XP magnetic bead purifying, carrying out a qPCR, carrying out high-fidelity PCR amplification on a library by utilizing a qPCR result, carrying out targeted capture on the cancer gene of the library, and the like. By utilizing the method disclosed by the invention, a relatively high library yield can be obtained, the defect that the quality of the library is unstable due toctDNA quality difference is overcome, the sensitivity and accuracy of detection are improved fundamentally, and the false positive rate of detection is reduced. Therefore, the method can be used for preparing a variety of tumor tracing or tumor screening kits to meet various clinical and research requirements in the field of biomedicine.

High-sensitivity double-chain circulating tumor DNA detection method and kit

The invention discloses a multi-amplification database building method for trace DNA and a special kit. The multi-amplification database building method can realize multiple amplification sequencing with trace DNA or even single-cell-level DNA as a template, and the whole reaction process can be realized in a single tube. Experiments prove that the database building method achieves 207-fold one-tube amplifier amplification at the single cell level, has the fidelity higher than that of conventional commercial kits, not only has the advantages of convenient operation, low cost, high sequencing quality and high sensitivity, but also can meet the sequencing requirements of two sequencing platforms, and is suitable for high-throughput and low-frequency mutation detection fields such as the ctDNA mutation detection field and the low-initial-quantity DNA embryonic mutation detection field such as single cell mutation detection and screening.

Multi-amplification database building method for trace DNA and special kit

The invention relates to the technical field of molecular biology and particularly relates to a magnetic bead reagent applied to screening super-long fragment nucleic acids and a screening method. Thereagent comprises silicon oxide hydroxyl magnetic beads with the concentration of 20-800ng/uL, Tris-HCl with the concentration of 5-50mmol/L, sodium chloride with the concentration of 1-60mol/L, sodium laurate with the concentration of 0.1-20mol/L, guanidine hydrochloride with the concentration of 0.1-10mol/L, NaN3 with the concentration of 0.1-100mol/L, polyethylene glycol with the volume percent concentration of 1-25% and EDTA (Ethylene Diamine Tetraacetic Acid) with the pH value of 8 and the concentration of 0.1-10mmol/L. The reagent aims at the problem in the prior art that the large-fragment DNA recovery effect is poor in the high-throughput sequencing library construction process, large fragments are separated and screened by using magnetic beads by designing and optimizing the buffer solution of the magnetic beads, the magnetic bead reagent is particularly applicable to the long-fragment single-molecule sequencing of PacBio, Nanopore and the like, the fragment length of the sequencing data can be greatly improved, and better sequencing results can be obtained.

Magnetic bead reagent applied to screening super-long fragment nucleic acids and screening method

The invention relates to the field of nucleic acid purification, and provides a method for removing primer dimers. The method comprises the following steps that (1) PCR products are purified by adopting an ultrafiltration centrifugation method; and (2) the product obtained in (1) is purified by using a magnetic bead method to further remove the primer dimers, wherein the difference value between the size of the PCR products and the primer dimers is 20 bp-170 bp. In addition, the invention further provides the application of a magnetic bead reagent in the primer dimer removing and a reagent combination for removing the primer dimers.

Method for removing primer dimers

The invention discloses a Clean-CL purifying kit and application thereof. The Clean-CL purifying kit is a magnetic bead method NDA product purifying kit and contains a magnetic bead purifying reagent, wherein the magnetic bead purifying reagent is a solution prepared by uniformly mixing Speed Bead Magnetic Carboxylate Modified Particles, a Tris-HCL solution, an EDTA solution, sodium chloride, polyethylene glycol and water, can be effectively combined with 100bp-500bp segments and is capable of effectively removing dNTP, primers, a primer dimer, joints, saline ions and other impurities. Experiments prove that the magnetic bead purifying reagent in the Clean-CL purifying kit is good in usability and stability and capable of guaranteeing the construction quality of a library and can be applied to the construction of Sanger and NGS libraries, the purification of a PCR system and a coupled reaction system and the like.

Clean-CL purifying kit and application thereof

The invention discloses a high-flux library construction kit and application thereof. The specific process for constructing a library by using the library construction kit comprises the following steps that firstly, incision enzymes are used for conducting tail end flattening repairing on genomic DNA of a sample to be detected, and tail ends are filled and flattened; then one adenine A is added to3' ends of repaired fragments by using ligase, and joints are added by using a TA connection method; finally, PCR amplification is conducted, DNA fragments connected with the joints are gathered, andthe concentration and fragment section which are required by on-machine sequencing are achieved. Compared with other library construction kits in domestic and foreign markets, the high-flux library construction kit has the advantages that the efficiency of connecting the DNA fragments with the joints is higher, the residual problem of joint self-connection in library construction is solved, the fixed concentration after connection is guaranteed, impurity removal is conducted more completely at the same time, most importantly, a large amount of cost is saved, and the high-flux library construction kit has good market prospects in DNA library construction of high-flux sequencing.

High-flux library construction kit and application thereof

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Clean-CL purifying kit and application thereof

High-flux library construction kit and application thereof