Cleaning method and precautions for PCR 8 Strip Tubes kit
We need to clean the PCR Octet Tubes kit after use, there are some methods to purify the 8 Strip PCR Tubes cleaning kit, what are the contents that need to be noted in the process of use?
Experimental method principle: silica gel membrane binds DNA under high salt conditions and can be separated from DNA under low salt conditions. Primers, mononucleotides, enzymes, mineral oil, salt ions, etc. in DNA-containing solutions used for cleaning purposes are separated because they do not have similar properties to DNA.
A, PCR Strip Tubes manufacturers talk about the composition of the kit, storage, stability
1, instructions, consumables: 96-well DNA preparation plate, 96-well 1.6ml deep-well plate, 96-well V-shaped plate.
2, Buffer PCR-A: DNA binding solution. Store in a sealed state at room temperature. If precipitation occurs, it should be dissolved in a warm bath at 65°C and cooled to room temperature before use.
3. Buffer W2 Concentrate: Remove the salt solution. Before use, add ethanol according to the volume specified on the bottle, mix thoroughly, and store at room temperature. You can use 100% ethanol or 95% anhydrous ethanol.
4. Elution solution: 2.5mM Tris-HCl, pH 8.5, stored in a sealed state at room temperature.
2. PCR Strip Tubes manufacturer talks about operation steps
Users can choose negative pressure or centrifugation.
A. Negative pressure method
1. Connect the negative pressure device correctly, put the 96-well DNA preparation plate on the negative pressure device; add 3 times buffer PCR-A (if buffer PCR-A is less than 100μl, add 100μl) to the PCR, enzyme digestion, enzyme labeling or sequencing reaction solution; mix thoroughly and transfer to the 96-well DNA preparation plate, turn on and adjust the negative pressure to -25-30 inch Hg, and slowly aspirate the solution from the plate.
2. Add 0.3 ml of buffer W2 and absorb the solution. Wash twice with 0.3 ml of buffer W2 in the same manner. Confirm the addition of anhydrous ethanol to the specified volume of Buffer W2 concentrate on the reagent bottle.
3, Maintain negative pressure and extract the 96-well DNA preparation plate for 10 minutes.
4, Crush the 96-well DNA preparation plate 6 times on long fibrous tissue with the drainage tube facing downward.
5, Place the 96-well DNA preparation plate on a 96-well V-shaped plate, add 25-30 ul of water or elute to the center of the membrane, and let stand for 1 min at room temperature. Elute DNA by centrifugation at 3 000 x g for 5 minutes.
B. Centrifugation
1. Add 3 times the volume of Buffer PCR-A (add 100 μl if Buffer PCR-A is less than 100 μl) to the PCR, digestion, enzyme labeling or sequencing reaction; mix and transfer to 96 wells of DNA in 96 wells of the preparation plate. Place the DNA preparation plate in a 96-well 1.6 ml deep well plate, centrifuge at 1000 × g for 1 min, and discard the filtrate.
2. In the 96-well DNA preparation plate, add 0.3 ml of buffer W2, centrifuge at 1000×g for 1 min, and discard the filtrate. Wash again with 0.3 ml of buffer W2 in the same manner. Confirm the addition of anhydrous ethanol to the specified volume of Buffer W2 concentrate on the reagent bottle.
3. Place the 96-well DNA preparation plate in a 96-well 1.6 ml deep well plate and centrifuge at 3 000 x g for 10 minutes.
4, Place the 96-well DNA preparation plate in a clean 96-well V-bottom plate, add 25-30 ul of water or elute to the center of the membrane, and let stand at room temperature for 1 minute. Elute DNA by centrifugation at 3 000 x g for 5 minutes.
PCR 8 Strip Tubes manufacturer talks about precautions.
1, Heating the eluate or water to 65°C helps to improve the elution efficiency.
2. DNA molecules are acidic and storage in 2.5 mM Tris-HCl, pH 8.5 eluent is recommended.
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