First, the material
Young leaves.
Second, the equipment
Pipette, frozen high speed centrifuge, bench top high speed centrifuge, water bath, ceramic mortar, 50 ml centrifuge tube (with lid) and 5 ml and 1.5 ml centrifuge tubes, bent into small hooks.
Third, the reagent
1. Extraction buffer I: 100 mmol/LTris·Cl, pH 8.0, 20 mmol/L EDTA, 500 mmol/L NaCl, 1.5% SDS.
2. Extraction buffer II: 18.6 g of glucose, 6.9 g of sodium diethyldithiocarbonate, 6.0 g of PVP, 240 ul of mercaptoethanol, and water to 300 ml.
3, 80:4:16/chloroform: pentanol: ethanol
4, RnaseA mother liquor
5. Other reagents: liquid nitrogen, isopropanol, TE buffer, absolute ethanol, 70% ethanol, 3 mol/L NaAc.
Fourth, the operation steps:
1. Add 20 ml of extraction buffer I to a 50 ml centrifuge tube and preheat in a 60 °C water bath.
2. 5-10g of seedlings or leaves, cut into pieces, add liquid nitrogen into powder in the mortar, immediately pour into preheated centrifuge tube, shake vigorously and mix, heat in 60 °C water bath for 30-60 minutes (long time) , DNA yield is high), shaking from time to time.
3. Add 20ml chloroform / pentanol / ethanol solution, mix by inversion (with gloves, to prevent damage to the skin), let stand for 5-10 minutes at room temperature, layer the aqueous and organic phases (remix if necessary) .
4. Centrifuge at 5000 rpm for 5 minutes at room temperature.
5. Carefully remove the supernatant into another 50 ml centrifuge tube, add 1 volume of isopropanol, mix, and leave the flocculent DNA precipitate for a while at room temperature.
6. Add 1 ml of TE to 1.5 ml eppendorf. The DNA floc was taken out with a hook-shaped glass rod, blotted dry on a clean absorbent paper, transferred to a centrifuge tube containing TE, and the DNA was quickly dissolved in TE.
7. If the DNA does not form a flocculent precipitate, it can be centrifuged for 5 minutes at 5000 rpm and the pellet is transferred to the TE tube. The precipitate thus collected is often difficult to dissolve in TE and can be placed in a water bath at 60 ° C for more than 15 minutes to aid dissolution.
8. Centrifuge the DNA solution at 3000 rpm for 5 minutes and pour the supernatant into a clean 5 ml centrifuge tube.
9. Add 5 μl of RNaseA (10 μg/μl) and remove the RNA at 37 ° C for 10 minutes (RNA has no effect on DNA manipulation and analysis, and this step can be omitted).
10. Add 1/10 volume of 3mol/L NaAc and 2× volume of ice ethanol, mix and store at -20 °C for about 20 minutes, and the DNA will form a flocculent precipitate.
11. Use a glass rod to remove the DNA pellet, rinse with 70% ethanol, and blot dry on clean absorbent paper.
12. Re-dissolve the DNA in 1 ml TE, -20 storage.
13. 2 μl of DNA sample was electrophoresed on 0.7% Agarose gel to determine the molecular size of the DNA. At the same time, 15 μl was diluted 20 times, OD260/OD280 was measured, and DNA content and quality were measured.
[Note] 5g sample can guarantee 500μg DNA, which is sufficient for RFLP, PCR and other analysis.
Ribbon Transfer Machine
1. AA series ribbon transfer machine
This is a multi-function printing machine that can be applied for ribbon printing, cutting and rolling cloth. Moreover, it is a new generation of roller sublimation Heat Transfer Machine researched and developed by our senior technicians according to the current market demands. The machine runs stably and efficiently at a high speed with tri-loction , accurate positioning ,and prints with a couple of ribbon simultaneously in whole roll, thus to get a clear printing result.
1). Adopt the most advanced control system, imported components and auto spacing, which leads to the excellent performance (accurate and safe) and long lasting life. High precision (0.5 ℃), all digital display of time, and high accuracy.
2).Oil thermal conducting structure makes the uniform color of the transfer printing. Equipped with the simultaneous down-roller, which make minimal mold positioning error. It can transfer printed a couple of ribbons simultaneously.
3).Both single and mass production are available. The large diameter bearing makes the maintenance of the fragile parts of the heating system. The drive shaft is made by using Japanese [Shin Etsu" material; Blanket is made by using Du Pont material imported from USA and large capacity conduction oil, which make consistent temperature, without any color error even in continuous transfer printing.
2. AB series ribbon transfer machine
This product is a response to the ribbon, lace, mobile phone lanyard, doubled-side printing fabric. Our company is new product development team. The heating temperature stability, set accurate positioning, product transfer effect is clear, You can navigate according to different product size, and can transfer a plurality of products, improve the efficiency .
1). Using the most advanced imported components, control system, automatic limit, performance (accuracy, security), long service life, high precision, digital display system, high precision.
2). Blanket band separated, the lever , and a pressure balancing stem, ensure that every consistent blankets and heating, body pressure, complete solved in shut down or failure of blanket cooling problems, better protect the blankets, prolong its service life.
3). Roller adopts the shunt tube design, heating tube heating, heat conducting oil heat transfer, uniform temperature, no color printing effect. At the same time with the synchronous roller shaft. The unique positioning system, product does no shift, not white. It can transfer multiple products at the same time.
Ribbon Transfer Machine
Ribbon Transfer Machine,Thermal Ribbon Transfer Machine,Automatic Ribbon Transfer Machine
KC Printing Machine (Group) Limited , https://www.kcautopm.com