DNA-Sorb-D (CE) | K-1/8-100

DNA-Sorb-D | K-1/8-100 from Sacace Biotechnologies is available for delivery

Description:

General information: DNA extraction kit from liquid-based cytology samples (Cytoscreen, PreservCyt,..)

Target Disease Type: RNA&DNA purification

Specific Application: Silica Sorbtion and Heat-Based* Method

Storage and Shipping : on request

DNA-Sorb-D (CE) K-1/8-100 DataSheet

INTENDED USE

The DNA-Sorb-D nucleic acid extraction kit is intended for the isolation and purification of DNA from liquid-based cytology samples (Cytoscreen, PreservCyt, etc).

STABILITY

DNA-Sorb-D is stable up to the expiration date indicated on the kit label

SPECIMEN AND REAGENT PREPARATION

I step: Pretreatment of specimens

• Prepare required quantity of 2,0 ml screw-cap tubes for the samples.
• Vortex vigorously transport medium for liquid-based cytology and transfer 1,8 ml of cells to the appropriate tube.
• Centrifuge the tubes at 12000g for 10 min. Discard the supernatant and leave about 150 µl of solution for DNA extraction.

II step: Cells washing

• Add to the tubes 1 ml of Mycolisin, vortex vigorously and incubate at room temperature for 30 min. Vortex periodically. Centrifuge the tubes at 12000g for 2 min. Discard the supernatant and leave about 150 µl of solution.
• Add 1ml of PBS-buffer to each tube. Vortex and centrifuge the tubes at 12000g for 2 min. Discard the supernatant without disturbing the pellet.
• Vortex Cytolisin, add 0,1ml to each tube and mix by pipetting. Vortex the tubes and incubate 2 hours (or overnight) at room temperature.

III step: DNA purification

• Lysis Solution and Washing Solution (in case of their storage at +2-8°C) should be warmed up to 60–65°C until disappearance of ice crystals. Prepare one 1,5 ml tube for the Negative extraction Control.
• Add to each tube 10 µl of Internal Control (if provided with the amplification kit) and 300 µl of Lysis Solution.
• Prepare Controls as follows: add 100 µl of C– (Negative Control provided with the amplification kit) to the tube labeled Cneg.
• Vortex the tubes and incubate for 5 min at 65°C. Centrifuge for 7-10 sec. If the sample is not completely dissolved it is recommended to re-centrifuge the tube for 5 min at a maximum speed (12000-16000 g.) and transfer the supernatant into a new tube for DNA extraction.
• Vortex vigorously Sorbent and add 25 µl to each tube.
• Vortex for 5-7 sec and incubate all tubes for 3 min at room temperature. Repeat this step.
• Centrifuge all tubes for 30 sec at 5000g and using a micropipette with a plugged aerosol barrier tip, carefully remove and discard supernatant from each tube without disturbing the pellet. Change tips between the tubes.
• Add 500 µl of Washing Solution to each tube. Vortex vigorously and centrifuge for 30 sec at 10000g. Remove and discard supernatant from each tube.
• Repeat step 8 and incubate all tubes with open cap for 5-10 min at 65°C.
• Resuspend the pellet in 100 µl of DNA-eluent. Incubate for 5 min at 65°C and vortex periodically.
• Centrifuge the tubes for 1 min at 12000g.
• The supernatant contains DNA ready for amplification. If amplification is not performed the same day of extraction, the processed samples can be stored at 2-8°C for at maximum period of 5 days or frozen at –20°/-80°C.