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Method for simultaneously sequencing a plurality of nucleic acid samples

A nucleic acid sample and sequencing technology, which is applied in the sequencing and biological fields, can solve the problems of low cost and needs to be improved, and achieve the effects of low cost, cost-effective sequencing, and improved purity

Inactive Publication Date: 2014-02-26
田埂
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Problems solved by technology

However, there is still no low-cost and high-accuracy method for simultaneously sequencing multiple nucleic acid samples
[0003] Therefore, the current methods for simultaneously sequencing multiple nucleic acid samples still need to be improved

Method used

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  • Method for simultaneously sequencing a plurality of nucleic acid samples
  • Method for simultaneously sequencing a plurality of nucleic acid samples
  • Method for simultaneously sequencing a plurality of nucleic acid samples

Examples

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Embodiment 1

[0062] According to the method for simultaneously sequencing multiple nucleic acid samples of the present invention, refer to figure 1 , follow the steps below to sequence:

[0063] 1. Sample extraction

[0064] Free DNA extraction reagents are used to extract free DNA. The main steps are as follows: Temporarily store the whole blood sample in ice at 4°C, and turn on the high-speed refrigerated centrifuge. Centrifuge the blood collection tube at 1600g for 10 minutes at 4°C to remove residual cells, and then centrifuge at 16000g at 4°C After 10 minutes, remove residual cells and transfer the supernatant to a new 2ml centrifuge tube.

[0065] Add proteinase K600μL fully dissolved and mixed plasma in a 2mL centrifuge tube, vortex shaker to mix for 10s, and briefly centrifuge.

[0066]Add 600 μL of mixed GB mix and centrifuge briefly; place in a water bath at 56°C for 10 minutes; take out the centrifuge tube, cool at room temperature for 5 minutes, and centrifuge briefly; add 3...

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Abstract

The invention discloses a method for simultaneously sequencing a plurality of nucleic acid samples, which comprises: respectively performing adaptor ligation, first PCR amplification, terminal phosphorylation, and DNA ligation in order of the plurality of nucleic acid samples; mixing the plurality of DNA ligation products; adding a base A at the 3'-terminal of the mixed DNA ligation products; connecting the ligation products with the 3'-terminal added with the base A to a sequencing interface; performing second PCR amplification of the sequencing interface-connected products to obtain a mixed sequencing library of the plurality of nucleic acid samples; sequencing the mixed sequencing library; and classifying the sequencing results to respectively obtain respective nucleic acid sequences of the plurality of nucleic acid samples. The method of the invention can simultaneously perform sequencing of a plurality of nucleic acid samples, is accurate in sequencing results, good in repeatability, low in cost, and high in efficiency, makes full use of sequencing flux, and is especially suitable for blood free DNA samples.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to the field of sequencing technology. Specifically, the present invention relates to a method for simultaneously sequencing multiple nucleic acid samples. Background technique [0002] The second-generation deep sequencing technology represented by the Illumina Hiseq series has made great progress in recent years. Through the simultaneous parallel sequencing of millions of short DNA fragments, the sequencing of each base can be completed in a short time. , and compared with traditional sequencing, the cost is greatly reduced and the accuracy is improved. However, there is still no low-cost and high-accuracy method for simultaneously sequencing multiple nucleic acid samples. [0003] Therefore, the current method for simultaneously sequencing multiple nucleic acid samples still needs to be improved. Contents of the invention [0004] The present invention aims to solve at le...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2535/122
Inventor 田埂方建火郎继东张丽娜
Owner 田埂
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