Ilp Molecular Marker Primer of Arrowhead Pea and Its Application in Variety Identification
A technology of arrow peas and molecular markers, applied in the biological field, can solve problems such as lack of whole genome data, and achieve the effects of high sensitivity and accuracy, convenient operation and low cost
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[0063] 1. Identification material DNA extraction
[0064] Referring to the SDS method, the specific operation steps are as follows:
[0065] 1) Take 5 g of collected fresh leaves, put them into a mortar and grind them into powder, add 600 μl of DNA buffer (30% SDS lysate) into the mortar, and mix well.
[0066] 2) Pour the homogeneous solution into a centrifuge tube and place in a water bath at 60°C for 10 minutes, and shake it every 3-4 minutes during the process.
[0067] 3) Take it out and cool to room temperature, add 195 μl of 5M KAc solution, mix well and place on ice for 5 minutes.
[0068] 4) Add 600 μl of chloroform isoamyl alcohol (24:1) (operate in a fume hood with the ventilation system turned on), and mix well.
[0069] 5) Let it cool down to room temperature, centrifuge in a centrifuge at 12000r / min for 10min, and pipette 360μl of the supernatant into a clean centrifuge tube.
[0070] 6) Add 240 μl isopropanol and 36 μl 3M NaAc solution to the supernatant, mix...
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