Hybrid capture kit for genome target region sequencing, capture method and application

A target area, hybridization capture technology, applied in the field of hybridization capture kits, can solve the problems of low capture efficiency, difficult to meet, high price, etc., and achieve the effect of improving capture efficiency, reducing experimental cost, and good capture efficiency and uniformity

Pending Publication Date: 2022-05-13
上海英基生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many commercial kits using liquid phase hybridization capture technology, but they are mainly foreign industry giants, and the price is high
Domestic products generally have the problem of l

Method used

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  • Hybrid capture kit for genome target region sequencing, capture method and application
  • Hybrid capture kit for genome target region sequencing, capture method and application
  • Hybrid capture kit for genome target region sequencing, capture method and application

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0030] Example 1

[0031]The object of the present embodiment is to compare the performance of the kits and methods of the present invention with several commercially available similar products when hybridizing capture. Among the similar products, the domestic 1: Hybridization Capture Kit (P10006, Homgen): and the domestic 2: BOKE Capture Kit (TC003116, BOKE), two domestic brands. Foreign: xGen Hybridizationand Wash Kit (1080584, IDT), a foreign brand, the test of similar products is carried out according to their respective instructions.

[0032] The kit operation steps of the present invention mainly includes the prepared library with Human Cot DNA, Blocker mixing and blocking, hybridizing the sample with a probe after blocking, preparation of elution buffer and streptavidin bead preparation, bead capture and cleaning and Post-capture PCR. The details are as follows:

[0033] 1 The prepared library is mixed with Cot DNA and Blocker to concentrate

[0034] 1.1 Place The Universa...

Example Embodiment

[0119] Example 2

[0120] The present embodiment is to test the hybridization capture effect of betaine (Beta) and tetramethylammonium chloride (TMAC) in different ratios in hybridization buffer. The experimental operation process refers to Example 1, which is not listed in detail here, and the tested configuration is as follows:

[0121] Test Beta TMAC Basal hybridization buffer Test1 2M 1.5M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sulfate Test2 2M 1M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sulfate Test3 2M 0.5M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sulfate Test4 1.5M 1.5M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sulfate Test5 1.5M 1M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sulfate Test6 1.5M 0.5M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sulfate Test7 1M 1.5M 2x SSPE, 5x Denhandt's, 2mM EDTA, 0.02% SDS, 10% dextran sul...

Example Embodiment

[0125] Example 3

[0126] The following embodiment is to test the amount of hybridization buffer enhancer in the hybridization system, the experimental operation process refers to Example 1, which is not listed in detail here, the enhancer selects formamide and guanidine thiocyanate, and its proportion and the corresponding hybridization system and the adjustment of the magnetic bead capture system are as follows:

[0127] Hybridization system:

[0128] Test formamide Guanidine thiocyanate (6M) Hybridization buffer probe NFWater Test1 2ul 8.5ul 4ul 2.5ul Test2 2.7ul 8.5ul 4ul 1.8ul Test3 3.5ul 8.5ul 4ul 1ul Test4 2ul 8.5ul 4ul 2.5ul Test5 2.7ul 8.5ul 4ul 1.8ul Test6 3.5ul 8.5ul 4ul 1ul Test7 2.7ul 1.8ul 8.5ul 4ul Test8 2.7ul 1ul 8.5ul 4ul 0.8ul Test9 2.7ul 0.5ul 8.5ul 4ul 1.3ul Test10 2.4ul 0.3ul 8.5ul 4ul 1.8ul Test11 2ul 0.7ul 8.5ul 4ul 1.8ul Test12 1.7ul ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a hybridization capture kit for genome target region sequencing, a capture method and application. The kit provided by the invention is suitable for genome libraries of various different sources, can effectively reduce the experiment cost and improve the capture efficiency, and is suitable for sequencing a genome target area. Experiments prove that compared with several commercially available similar kits, the kit provided by the invention has better capture efficiency, uniformity and other aspects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a hybrid capture kit, a capture method and an application for sequencing a genome target region. Background technique [0002] In recent years, with the rapid development of sequencing technology, high-throughput and high-accuracy next-generation sequencing technology (NGS) is being more and more widely used. High-throughput sequencing has greatly reduced the cost of sequencing. However, at present, the cost of whole-genome resequencing is still very high, and the huge amount of data obtained brings a lot of inconvenience to subsequent analysis, making it impossible for large-scale application. Targeted sequencing technology can enrich the genomic regions of interest for sequencing, greatly reducing the output of sequencing data, data analysis is simple and fast, and can take advantage of high-throughput sequencing technology more cost-effectively. Targeted sequencing t...

Claims

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Application Information

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IPC IPC(8): C12Q1/6813C12Q1/6806C40B50/06C12Q1/6869
CPCC12Q1/6813C12Q1/6806C40B50/06C12Q1/6869C12Q2565/549C12Q2525/191C12Q2531/113C12Q2523/308C12Q2535/122
Inventor 冯延叶孙大坤柴智刘会珍孙扬
Owner 上海英基生物科技有限公司
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