Second, equipment, frozen desktop centrifuge, low temperature vacuum dryer, electrophoresis apparatus, electrophoresis tank.
Third, reagent
TE-saturated phenol: chloroform (1:1), chloroform, 3M NaAc (pH 5.2), ethanol (100% and 70%), TE buffer, RNase-free double bacteria water.
Fourth, the operation steps
1. Take an eppendorf tube and purify 400 ml of TMV (10 mg/ml). Add an equal volume of phenol/chloroform. Close the lid of the tube and shake well for 1 minute by hand. Centrifuge at 12000 g for 10 minutes at 4°C.
2. Pipette the aqueous phase in a new eppendorf tube and extract with phenol/chloroform until the interface between the aqueous and organic phases is free of protein.
3. Pipette the aqueous phase into a new eppendorf heart tube, add an equal volume of chloroform, invert the centrifugation tube by hand for tens of seconds, and centrifuge at 12,000g for 10 minutes at 4°C.
4. Take the aqueous phase, add 1/10-fold volume of 3mol/L NaAc (pH5.2), 2.5 volumes of ice-cold ethanol, and mix well, -20°C for 30 minutes.
5. Centrifuge at 12000g for 15 minutes at 4°C, carefully discard the supernatant, wash the pellet with 70% ethanol, centrifuge at 12000g for 5 minutes at 4°C.
6. The supernatant was carefully discarded and the precipitate was vacuum dried for 5 minutes or air dried for 10 minutes and dissolved in RNase-free double bacteria or TE buffer.
7. Take 10ml for electrophoresis analysis and another 10ml for cDNA synthesis.
[Note] 1. The entire operation should be carried out at a low temperature as much as possible.
2. Since the viral RNA is embedded in the coat protein, it is necessary to fully strip the virus coat protein, and it is generally necessary to perform phenol/chloroform extraction several times.
Green Tea is unleavened tea that is not made by fermentation. Because of the different processing method, tea have two kinds shape named Gunpowder Tea and chunmee tea, gunpowder is round shape likes pearl and chunmee is strip shape likes eyebrow. Green tea retains more natual substances of fresh leaves and contains more nutrients such as tea polyhenols, catechins, chlorophyll, caffeine, amino acids and vitamins. These natural nutrients has many function such as:
1. Anti – aging effects;
2. Disease suppression;
3. Preventive and anticancer properties;
4. Radiation protection;
5. Resisting virus bacteria;
6. Beauty & Skincare effect;
7. Refresh the mind;
8. Effect of diuresis;
9. Relieve fatigue;
10. Protecting teeth and improving eyesight;
11. Eliminate bad breath;
12. Reduce high blood pressure, high blood lipids, high blood sugar;
13. Reducing fat and promoting digestion;
14. Visceral fat suppression;
And so on.
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