Method for rapid and high-flux extraction of plant genome DNA and application of plant genome DNA
A plant genome, high-throughput technology, applied in the field of molecular biology, can solve the problems of cumbersome extraction process, time-consuming and labor-consuming, etc., and achieve the effect of high extraction throughput, low cost and high applicability
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Embodiment 1
[0028] Cut about 1 g of fresh corn leaves, which can be directly used for genome extraction, or stored at 4°C for later use, and extracted within 10 days. Put the leaves and a steel ball with a diameter of about 3 mm into a 1.5 ml centrifuge tube, and add 300 μL of modified SDS extract. The formula of the SDS extract is: 100mM Tris·HCl, 50mM EDTA, 100mM NaCl, 1% w / v SDS, pH 8.0.
[0029] Move the centrifuge tube to the sample grinder of the high-throughput tissue grinder, set the frequency at 33 times / s, shake for 30 s, put the ground sample at 65°C and heat for 5-10 min, and mix by inverting 3-4 times during the period. Put the heated centrifuge tube into a centrifuge, and centrifuge at room temperature for 10 min at 12000 rpm. Take 100 μL of supernatant in each tube, put it into a 96-well plate, add an equal volume of isopropanol with a row gun, cover each plate with a silica gel pad, mix it upside down, and let it stand at room temperature for 30 minutes.
[0030] Centrif...
Embodiment 2
[0035] Cut about 1g of fresh soybean leaves, and the leaves can be directly used for genome extraction, or stored at 4°C for later use, and extracted within 10 days. Put the leaves and a steel ball with a diameter of about 3 mm into a 1.5 ml centrifuge tube, and add 200 μL of modified SDS extract. The formula of the SDS extract is: 80mM Tris·HCl, 60mM EDTA, 120mM NaCl, 0.5% w / v SDS, pH 8.0.
[0036]Move the centrifuge tube to the sample grinder of the high-throughput tissue grinder, set the frequency at 33 times / s, shake for 30 s, put the ground sample at 60°C and heat for 5-10 min, and mix by inverting 3-4 times during this period. Put the heated centrifuge tube into a centrifuge, and centrifuge at room temperature for 10 min at 10,000 rpm. Take 100 μL of supernatant in each tube, put it into a 96-well plate, add an equal volume of isopropanol with a row gun, cover each plate with a silica gel pad, mix it upside down, and let it stand at room temperature for 30 minutes.
[...
Embodiment 3
[0042] Cut about 1g of fresh rice leaves, and the leaves can be directly used for genome extraction, or stored at 4°C for later use, and extracted within 10 days. Put the leaves and a steel ball with a diameter of about 3 mm into a 1.5 ml centrifuge tube, and add 500 μL of modified SDS extract. The formula of the SDS extract is: 120mM Tris·HCl, 40mM EDTA, 80mM NaCl, 1.5% w / v SDS, pH 8.0.
[0043] Move the centrifuge tube to the sample grinder of the high-throughput tissue grinder, set the frequency at 33 times / s, shake for 30 s, put the ground sample at 70°C and heat for 5-10 min, and mix by inverting 3-4 times during this period. Put the heated centrifuge tube into a centrifuge, and centrifuge at room temperature for 10 min at 12000 rpm. Take 100 μL of supernatant in each tube, put it into a 96-well plate, add an equal volume of isopropanol with a row gun, cover each plate with a silica gel pad, mix it upside down, and let it stand at room temperature for 30 minutes.
[004...
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