A kit and method for efficiently extracting DNA from sugarcane sugar products
A technology of cane sugar and kit, applied in the field of molecular biology, can solve the problems of low yield and purity of DNA, unable to meet the requirements of the test, etc., and achieve the effect of improving the purity of extracted DNA
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Embodiment 1
[0055] 1. Five methods to extract DNA from brown sugar
[0056] Method 1: Using the pretreatment solution, lysate and DNA precipitation solution of the present invention
[0057] (1) Pretreatment: Take 5g of brown sugar and add 10mL pretreatment solution, the pretreatment solution is 10% (w / v) PEG8000, put it into -20℃ for precipitation, the treatment time is 48h, then centrifuge at 10000rpm for 30min, remove the supernatant .
[0058] (2) Preliminary extraction: Add 800 μl of lysis solution (the composition of the lysis solution is as follows: 500 mM NaCl, 50 mM Tris-HCl, 50 mM EDTA, 3% (w / v) SDS, 2% (w / v) PVPP, pH 8.0, solvent (water) to dissolve the precipitate, and repeatedly wash the inner wall of the centrifuge tube, transfer the liquid to a 1.5ml centrifuge tube, incubate at 65°C for 30 minutes, and mix well by inverting during the period; add 100 μL of 10M NH 4 AC, place on ice for 10min, centrifuge at 4°C and 12000rpm for 10min, take the supernatant; add an equal vo...
Embodiment 2
[0076] One, three methods to extract DNA in brown sugar
[0077] Method 1, magnetic bead binding solution, washing solution, eluent and magnetic beads of the present invention 1
[0078] (1) Pretreatment: Take 5g of brown sugar and add 10mL pretreatment solution, the pretreatment solution is 10% (w / v) PEG8000, put it into -20℃ for precipitation, the treatment time is 48h, then centrifuge at 10000rpm for 30min, remove the supernatant .
[0079] (2) Preliminary extraction: Add 800 μl of lysis solution (the composition of the lysis solution is as follows: 500 mM NaCl, 50 mM Tris-HCl, 50 mM EDTA, 4% (w / v) SDS, 2% (w / v) PVPP, pH 8.0, solvent (water) to dissolve the precipitate, and repeatedly wash the inner wall of the centrifuge tube, transfer the liquid to a 1.5ml centrifuge tube, incubate at 65°C for 30 minutes, and mix well by inverting during the period; add 100 μL of 10M NH 4 AC, place on ice for 10min, centrifuge at 4°C and 12000rpm for 10min, take the supernatant; add an ...
Embodiment 3
[0091] One, three methods to extract DNA in brown sugar
[0092] Method 1, kit and method of the present invention extract the DNA in commercially available brown sugar:
[0093] (1) Pretreatment: Take 5g of brown sugar and add 10mL pretreatment solution, the pretreatment solution is 10% (w / v) PEG8000, put it into -20℃ for precipitation, the treatment time is 48h, then centrifuge at 10000rpm for 30min, remove the supernatant .
[0094] (2) Preliminary extraction: Add 600 μl of lysate (the composition of lysate is as follows: 500 mM NaCl, 50 mM Tris-HCl, 50 mM EDTA, 4% (w / v) SDS, 2% (w / v) PVPP, pH 8.0, solvent water), dissolve the precipitate, and repeatedly wash the inner wall of the centrifuge tube, transfer the liquid to a 1.5ml centrifuge tube, incubate at 65°C for 30 minutes, and mix well by inverting during the period; add 80 μL of 10M NH 4 AC, place on ice for 10 min, centrifuge at 12000 rpm for 10 min at 4°C, take the supernatant; add an equal volume of DNA precipitat...
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