Universal kit and method for extracting DNA from trace plant materials
A plant material and kit technology, which can be used in DNA preparation, recombinant DNA technology, and microbial assay/inspection, etc., and can solve problems such as time-consuming and labor-intensive
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Embodiment 1
[0078] The DNA extraction kit of embodiment 1 whole genome sequencing of plants is to the extraction result of the plant material of different weights
[0079] 1. Experimental method
[0080] Take 0.0001, 0.0005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, and 1 gram of fresh soybean leaf tissue respectively, and add 600 μL of AP1 Buffer and 6 μL RNaseA (10 mg / mL), machine-polished samples in a grinding tube; 0.5 and 1 g of leaf samples were fully ground into a fine powder by adding liquid nitrogen in a mortar and transferred the fine powder to a 1.5 mL centrifuge tube, Add 600 μL AP1 buffer and 6 μL RNaseA (10 mg / mL), place at 65°C for 10 minutes, invert the centrifuge tube 2 to 3 times during the standing process, and mix the sample;
[0081] Add 200μL AP2 buffer, mix well, centrifuge at 13,000rpm for 5 minutes, carefully pipette the supernatant into a new 1.5mL centrifuge tube;
[0082] Add 1.5 times the volume of supernatant AP3 buffer, and immediately mix thoroughly by pipettin...
Embodiment 2
[0097] The DNA extraction kit of embodiment 2 whole genome sequencing of plants is to the extraction result of different kinds of plant materials
[0098] 1. Experimental method
[0099] Take 5 mg each of barley leaves, corn leaves, rice leaves (Japanese nitrile), sorghum leaves, soybean leaves, tomato leaves, dandelion leaves, wild chrysanthemum leaves, pothos leaves, gladiolus leaves, and pine needles, and add 600 μL of AP1 buffer and 6 μL of RNaseA (10mg / mL), use a proofer to crush the sample, place it at 65°C for 10 minutes, and mix the sample by inverting the centrifuge tube 2 to 3 times during the placement process;
[0100] Add 200 μL AP2 buffer, mix thoroughly, centrifuge at 13,000 rpm for 5-10 minutes, and carefully pipette the supernatant into a new 1.5mL centrifuge tube;
[0101] Add 1.5 times the volume of supernatant AP3 buffer, and immediately mix thoroughly by pipetting. If flocculent precipitation occurs at this time, it will not affect DNA extraction;
[010...
Embodiment 3
[0115] Example 3 Extraction Results of Different Parts of Plant Material by the DNA Extraction Kit for Plant Whole Genome Sequencing
[0116] 1. Experimental method
[0117] Take 5 mg of different parts of the root, stem, flower, fruit, and seed of the plant, add 600 μL of AP1 buffer solution and 6 μL of RNaseA (10 mg / mL), break the sample with a proofer, place it at 65 ° C for 10 minutes, and centrifuge it upside down during the placement process tube 2 to 3 times, mix the sample;
[0118] Add 200 μL AP2 buffer, mix thoroughly, centrifuge at 13,000 rpm for 5-10 minutes, and carefully pipette the supernatant into a new 1.5mL centrifuge tube;
[0119] Add 1.5 times the volume of supernatant AP3 buffer, and immediately mix thoroughly by pipetting. If flocculent precipitation occurs at this time, it will not affect DNA extraction;
[0120] Put the mixture (solution and possible flocculent precipitate) obtained in the previous step into an adsorption column DB (the adsorption co...
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