A molecular recognition method for high-efficiency grafted camellia root and ear combination of old Camellia oleifera trees
A technology for molecular identification and camellia oleifera tree, applied in the biological field, can solve the problems of no molecular identification technology of camellia oleifera grafted stock and ear combination, no inspiration of stock and ear combination, etc., and achieves easy popularization and expansion, reduction of high disease incidence, Simple to use effects
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[0025] (1) Using 7 kinds of clones of old Camellia oleifera and 48 kinds of camellias as materials, the DNA was extracted respectively by using the improved CTAB method. The steps are as follows:
[0026] 1) Put 10 grams of leaves of old camellia oleifera tree or camellia flower into a mortar, pour in an appropriate amount of liquid nitrogen for grinding, transfer to a centrifuge tube pre-added with 700 μL 2×CTAB, place in a water bath at 65°C for 45-60 minutes, and centrifuge ( 4°C, 1000rpm, 10min) to obtain supernatant I;
[0027] 2) Take 600 μL of the supernatant I in the centrifuge tube in step 1) and put it into a new centrifuge tube, add a mixture of phenol, chloroform and isoamyl alcohol (volume ratio 25:24:1) with a total volume of 600 μL, shake well and centrifuge (4°C, 1000rpm, 10min) to obtain supernatant II;
[0028] 3) Take 550 μL of the supernatant II into a new centrifuge tube, add 500 μL of 10×CTAB, shake well, put it in a 65°C water bath to dissolve for 2-3 m...
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