Catalogue Number: PP-226L-JEN
|Shelf Life:||12 months|
|Type:||DNA Isolation Kits|
Description: MagBeads Swabs DNA Preparation Kit is developed for easy, efficient and fast isolation of DNA from epithelial swabs and scrapings. The isolated DNA is suitable for PCR or other molecular biology applications like labeling, cloning or sequencing. The kit contains all necessary reagents in one box. The isolation procedure can be easily adapted for isolation of smaller or larger amounts of DNA. The principle of the method is based on a reversible adsorption of DNA on magnetic particles coated with a special SiO2 surface. A schematic isolation procedure is shown on Figure 1. The sample cells are lysed and the containing DNA is bound to magnetic particles. After a repeated washing cycle the purified DNA is eluted from the magnetic particles. Caution of Strong Magnetic Field Magnetic fields can cause damage to magnetic storage media including credit cards, magnetic data tapes and computer hard drives. Strong magnetic fields can cause implanted heart pacemakers and cardioverter defibrillators to cease operation. Keep any electronic devices at least 1 Meter away from the magnets or magnetic racks. Interaction with metallic objects may produce pinch hazards.
Preparation procedure: 1 Before StartingResuspension of the magnetic particles is critical to achieve optimal results. Resuspend magnetic particles carefully by mixing or vortexing before use.Check all buffers for precipitates. If a visible precipitation appears, place the buffer in a warm water bath (35-50°C) and shake it until it becomes clear. The formation of a precipitate does not affect the quality of buffers.Mix all buffers well before use. 2 Scaling up the Preparation VolumeUse the following amounts of buffers and kit components for DNA preparation from different kind of swabs. Component||Amount per prep Sample Buffer (only required for isolation from buccal swabs)||300 µl Lysis Buffer||120 µl Binding Buffer||160 µl Magnetic particles DNA Grade||10 µl First Washing Buffer||300 µl Second Washing Buffer||300 µl Third Washing Buffer||300 µl Final Washing Buffer||300 µl Elution Buffer||50 µl 3 Sample Preparation 3.1 DNA isolation from buccal swabsAdd 300 µl Sample Buffer to a 1.5 ml tube.Collect buccal epithelium with a sterile cotton swab by gently rubbing the inner side of the donors check for approx. 20 sec.Insert the collected buccal swab into the tube containing Sample Buffer.Mix gently by manual swirling for 1 min.Remove the cotton swab collector, carefully pressing it against the tube wall to retain the excess buffer in the tube.Centrifuge the tube at 5,000 g for 3 min.Carefully remove the supernatant. 3.2 DNA isolation from vaginal or cervical swabsIn case the swab collector was not removed from the tube, centrifuge the tube briefly to drop the liquid from the tube wall and cap.Remove the cotton swab collector, carefully pressing it against the tube wall to retain the excess buffer in the tube.Mix the swab sample buffer and transfer up to 1 ml to a 1.5 ml tube.Centrifuge the tube at 5,000 g for 3 min.Carefully remove the supernatant. 4 Cell LysisAdd 120 µl well mixed Lysis Buffer and mix again by pipetting.Incubate at room temperature for 5 min (longer incubation up to 15 min may increase DNA yield).Centrifuge the tube at 8,000 g for 3 min.Transfer 100 µl of the supernatant into a new tube. 5 DNA BindingPlace 160 µl Binding Buffer and 10 µl of well mixed Magnetic Particles into a clean tube and mix both components thoroughly.Add the prepared suspension of magnetic particles to the tube containing the prepared sample and mix thoroughly.Incubate for 5 minutes at room temperature, mix once or twice during this time.Place the tube for up to 1 minute in a magnetic rack to collect the magnetic particles.Discard the supernatant without disturbing the magnetic particles at the tube wall. 6 First WashingPlace the tube in a non-magnetic rack.Add 300 µl First Washing Buffer and mix thoroughly until a homogeneous suspension is obtained.Place the tube in a magnetic rack to collect the magnetic particles.Discard the supernatant without disturbing the magnetic particles at the tube wall. 7 Second WashingPlace the tube in a non-magnetic rack.Add 300