How to Clean the PCR 8-Tube Kit

We need to clean the 8-strip PCR tubes with caps after using them. There are some ways to purify the PCR 8-tube cleaning kit. What should be paid attention to during use?

Principle of experimental method: Silica gel membrane binds DNA under high-salt conditions, and can be separated from DNA under low-salt conditions. Primers, single nucleotides, enzymes, mineral oil, salt ions, etc. in DNA-containing solutions for cleaning purposes are separated because they do not have similar properties to DNA.

Experimental materials: PCR products(8 strip PCR tubes with caps), reagents, PCR 8-Tube Kit, PCR cleaning kits, instruments, consumables, 96-well DNA preparation plates, 96-well deep-well plates, and V-shaped 96-well plates.

PCR tubes 0.2 ml 8 tube strips 02-ml-8-strip-pcr-tubes 0.1-ml-8-strip-pcr-tubes

The operation steps of cleaning the PCR 8-Tube Kit:

A. Negative pressure method

1. Correctly connect the negative pressure device, place the 96-well DNA preparation plate on the negative pressure device; add 3 times buffer PCR-A to the PCR, enzyme digestion, enzyme labeling, or sequencing reaction solution (if the buffer PCR-A is less than 100 μL, add 100 μL); mix well and transfer to a 96-well DNA preparation plate, turn on and adjust the negative pressure to -25-30 inches Hg, and slowly aspirate the solution in 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 absolute ethanol to the indicated volume of Buffer W2 concentrate on the reagent bottle.

3. Maintain the negative pressure and extract the 96-well DNA preparation plate for 10 minutes.

4. Grind the 96-well DNA preparation plate 6 times on the long fibrous tissue with the drainage tube facing down.

5. Place the 96-well DNA preparation plate on a V-shaped 96-well plate, add 25-30μL of water or elute to the center of the membrane, and let stand for 1 minute at room temperature. DNA was eluted by centrifugation at 3000 x g for 5 min.

B. Centrifugal

1. Place the DNA prep plate in a 96-well 1.6 mL deep well plate, centrifuge at 1000 x g for 1 min, and discard the filtrate. Add 3 volumes of Buffer PCR-A (if Buffer PCR-A is less than 100 μl, add 100 μl) to PCR, digestion, enzyme labeling, or sequencing reactions; mix and transfer to 96-well DNA in a 96-well preparation plate.

2. In a 96-well DNA preparation plate, add 0.3 mL of buffer W2, centrifuge at 1000×g for 1 minute, and discard the filtrate. Wash again in the same manner with 0.3 mL of buffer W2. Confirm the addition of absolute ethanol to the indicated volume of Buffer W2 concentrate on the reagent bottle.

3. Place the 96-well DNA preparation plate in a 96-well 1.6mL deep-well plate and centrifuge at 3000×g for 10 minutes.

4. Place the 96-well DNA preparation plate in a clean 96-well V-shaped bottom plate, add 25-30μL of water or elute to the center of the membrane, and let stand for 1 minute at room temperature. DNA was eluted by centrifugation at 3 000 x g for 5 min.

The matters needing attention in cleaning the PCR 8-Tube Kit:

1. Heating the eluent or water to 65°C can help improve elution efficiency.

2. DNA molecules are acidic, it is recommended to store them in 2.5 mM Tris-HCl, pH 8.5 eluent.

PCR series laboratory appliance also includes 0.1mL, 0.2mL 8 Strip PCR tubes, and 0.2mL Single PCR Tubes.

There are more introduces about PCR tubes: the use of PCR tubes, what are PCR tubes about the advantages, etc.