Method for high-throughput extraction of DNAs from rice leaves
A high-throughput, rice-based technology, applied in DNA preparation, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of low throughput, unsuitable labeling and typing of a small amount of pure line materials, and time-consuming, etc. Realize and realize the effect of high-throughput automation and low dependence on experimental conditions
Active Publication Date: 2017-11-21
HUAZHI RICE BIO TECH CO LTD
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However, this method takes a long time, has low throughput, and uses a large amount of toxic organic solutions such as chloroform. It is only suitable for sequencing or gene chips and other experiments that require high DNA quality. For large-scale high-throughput leaf DNA extraction such as gene identification, It is very inappropriate to mark and type a small amount of pure line materials
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[0035] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Those who do not indicate specific techniques or conditions in the embodiments, according to the techniques or conditions described in documents in the art, such as Sambrook and other molecular cloning experiment manuals (Sambrook J&Russell DW, Molecular Cloning: a Laboratory Manual, 2001), or according to the product Instructions are carried out. The reagents or instruments used were not indicated by the manufacturer, and they were all conventional products that can be purchased through formal channels.
[0036] In the following experiments, the steps of the rice leaf DNA extraction method (hereinafter referred to as the one-step method) are as follows (for the DNA extraction process, see figure 1 ):
[0037] ①Put about 20mg of rice leaves into a 2mL 96-well plate, and place the 96-well plate in an oven at 65°C for 60 minutes;
[00...
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The invention relates to a method for high-throughput extraction of DNAs from rice leaves. The method comprises the following steps: adding a proper amount of rice leaves into a proper container and carrying out baking at 55 to 65 DEG C until the rice leaves are dry; then grinding the rice leaves; then adding a DNA extraction solution, carrying out uniform mixing and then carrying out extraction; subjecting an extract obtained in the previous step to centrifugation; and weighing a proper amount of a supernatant, adding ultrapure water accounting for 19 to 60 times the volume of the supernatant and carrying out uniform mixing so as to obtain DNA. The method has the advantages that no organic solvent is used, so the method is safe and nontoxic; treatment with liquid nitrogen is not needed, and drying of the rice leaves with a baking oven is low in dependence on experimental conditions, easy to realize and free of influence on results; and a highly-matching pipetting workstation is employed, so high-throughput automatic operation is realized.
Description
technical field [0001] The invention relates to a method for extracting DNA from rice leaves with high throughput. Background technique [0002] Rice leaf DNA extracted by CTAB and other methods can meet the requirements of KASP genotyping, and good results can be obtained on Arraytape. However, this method takes a long time, has low throughput, and uses a large amount of toxic organic solutions such as chloroform. It is only suitable for sequencing or gene chips and other experiments that require high DNA quality. For large-scale high-throughput leaf DNA extraction such as gene identification, It is very inappropriate to mark and type a small amount of pure line materials. Therefore, it is necessary to develop a method for rapidly extracting DNA from rice leaves without using toxic organic reagents. [0003] Through the awesome "boiling method for rapid preparation of rice genomic DNA PCR template", we know that using TE buffer for boiling water bath can extract rice leaf...
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IPC IPC(8): C12N15/10C12Q1/68
CPCC12N15/1006C12Q1/6895C12Q2527/125
Inventor 李宇飞肖金华郑秀婷许娜彭佩李乐
Owner HUAZHI RICE BIO TECH CO LTD