24-well Magnetic Separator You will need a magnetic rack (separator) for use with the GeneCatcher™ gDNA 0.3-1 ml Blood Kit.

Store beads at room temperature. Resuspend the spheroplasts in 180 µl Lysis Buffer (L6) supplied with the kit.

Place the sample on the 24-well magnetic Separator for 1 minute. no.

Genomic DNA is extracted from blood samples using the cost-effective, user-friendly GeneCatcher™ Technology without the use of centrifugation. Place the tube on a 50 ml Tube Magnetic Separator Rack (see above for a figure) for 3 minutes. The procedure is designed for isolating gDNA using the GeneCatcher™ Magnetic Beads procedure. Protease (25 mg/ml in 50 mM Tris-HCl, pH 8.5, 5 mM CaCl, Protease Buffer (20 mM Tris-HCl, pH 8.5, 6 M guanidine HCl), Pellet of beads disturbed or lost during binding or washing steps.

Aliquot DNA and store at 4°C or -20°C. Tip: If the Elution Buffer is too viscous, add additional 150-250 µl Elution Buffer (E5) and keep the samples overnight at room temperature at this stage to increase the final yield. The elution buffer, Added Lysis Buffer containing Proteinase K during second rebinding step, Didn’t add Purification Buffer during second rebinding step. Vortex the tube containing the ChargeSwitch. Ensure the pieces are completely immersed in buffer. Stay up to date with G-Biosciences by signing up for our newsletter.

The manual also contains protocols for reaction cleanup and extraction of gDNA from …

Without removing the plate from the Magnetic Separator, carefully remove and discard the supernatant using a 1 ml pipette without disturbing the pellet of beads. To avoid repeated freezing and thawing of DNA, store the purified DNA at 4°C for immediate use or aliquot the DNA and store at -20°C for long-term storage, Add resuspended GeneCatcher™ Magnetic Beads and Lysis Buffer (L13) to a sterile 50-ml tube.

Add 1 ml Elution Buffer (E5; 10 mM Tris-HCl, pH 8.5) irrespective of the blood volume to the tube and gently agitate the sample by swirling the tube to release the bead pellet from the tube wall. Use the correct order of Wash Buffers for washing. Harvest up to 2 x 109 E. coli cells by centrifugation. Discard the flow through from the Wash Tube and place the cartridge into the Wash Tube.

Follow the recommendations below to obtain the best results: Perform all centrifugation steps at room temperature, Perform a 1 minute incubation step with Elution Buffer (E1) or water, Be sure to perform the recommended wash steps to obtain the best results, Always use sterile water, pH 7-8.5, if you are using water for elution. Binding DNA Follow the procedure below to bind DNA to the GeneCatcher™ Magnetic Beads. Without removing the plate from the Magnetic Separator, carefully remove the supernatant containing the DNA using a 1 ml pipette without disturbing the pellet of beads and transfer the supernatant to a sterile tube. Be sure to add 96–100% ethanol to Wash Buffer (W5). This quick protocol is meant for experienced users. Mix well by vortexing for 5 seconds. Introduction The flow chart for purifying genomic DNA using the PureLink™ Genomic DNA Purification Kit is shown below. The cells or tissues are digested with Proteinase K in the presence of EDTA to inhibit DNases. For high DNA content, incubate the samples for an additional 30 minutes. Centrifuge the column at maximum speed for 1.5 minute at room temperature. Optional: Add 20 µl RNase A (supplied with the kit) to lysate and incubate at room temperature for 2 minutes. To recover more DNA, perform a second elution step with 200 µl Elution Buffer (E1) or sterile, distilled water (pH >7.0) using another sterile 1.5 ml microcentrifuge tube. Other magnetic separators may not provide similar magnetic strength or may not be compatible with the volumes used in the protocol. Be sure to add Proteinase K and SDS solution during lysis. CS15024) for 24-well plates to obtain the best results. Other magnetic separators may not provide similar magnetic strength or may not be compatible with the volumes used in the protocol.

Sensitivity: Linear responses over the range of 0.5µg-50µg protein, Flexible Protocols: Suitable for tube or Titer plate assays, Ready to use assay reagents and no preparation required, Genomic DNA Extraction Protocol: Isolating Your Genomic DNA from Your Sample (Part 1 of 2). Add 180 µl Lysis Buffer (L6) and 20 µl Proteinase K (supplied with the kit) to the tube and mix well. Incubate at 55°C with occasional vortexing until lysis is complete (~3 hours). Note: Absence of aggregate after 2-3 minutes indicates very low DNA levels in the sample. Incubate at room temperature for 2 minutes. Add 2-10 ml well mixed blood samples to the tube containing the beads and gently invert the capped tube 3 times to mix the beads. To avoid repeated freezing and thawing of DNA, store the purified DNA at 4° C for immediate use or aliquot the DNA and store at -20° C for long-term storage. For samples with high DNA content, incubate the tube over night and tip-mix gently before proceeding to the next step.

Place the spin cartridge in a sterile 1.5-ml microcentrifuge tube. The 24-well Magnetic Separator is a magnetic separation rack (see below) for use in protocols with magnetic beads.

We recommend using the 24-well Magnetic Separator (catalog No. If you are using a frozen cell pellet, proceed to Step 3. Remove the plate from the water bath and gently agitate the plate by swirling or placing the plate on a shaker set to low speeds until the bead pellet is completely dispersed. Without removing the plate from the Magnetic Separator, carefully remove and discard the supernatant using a 1 ml pipette without disturbing the pellet of beads by angling the pipette such that the tip is pointed away from the pellet.

The sample can be tissue, plant or animal cells, blood, viral DNA or any other DNA containing sample. Frozen blood samples or blood samples exposed to repeated freeze-thaw cycles, Old, archived blood samples and degraded samples, Maintain a sterile environment when handling DNA to avoid any contamination from DNases, Ensure that no DNase is introduced into the solutions supplied with the kit, Make sure that all equipment coming in contact with DNA is sterile, including pipette tips and tubes, Perform the recommended wash steps during the purification to obtain the best results.

Incubate at room temperature for 5 minutes to allow the DNA to bind to the beads. Remove the plate containing the pelleted magnetic beads (Step 11, above) from the Magnetic Separator.

The SDS precipitates in the presence of guanidine isothiocyanate. The extraction kit is designed for pelleted cells from either cultured eukaryotic cell lines or purified peripheral blood mononuclear cells (PBMC). Protein Cross-Linking & Protein Modification, Ion Exchange Chromatography Resins and Methods, Protein Extraction & Lysis Buffer (PE LB™) Systems, Molecular Biology Accessories, Buffers & Reagents, Biotechnology, Science for the New Millennium, Purification Resin Synthesis & Production. For Research Use Only. 24-well U-bottom deep-well plates (Whatman 24-well Round Bottom UniPlate, cat. Upon receipt, store components as follows: All components are guaranteed stable for 6 months when stored properly. Add 0.3-1 ml well mixed blood samples to the wells containing the beads and gently agitate the plate by swirling or placing the plate on a shaker set to low speeds. The supernatant should be clear, olive green color. Note: If the magnetic bead pellet does not easily disperse, warm the plate at 65°C for 5 minutes and pipet up and down gently using a 1 ml pipette tip set to 400 µl to disperse the pellet without forming any bubbles. Agitate the samples by gently swirling the plate to resuspend any settled beads. Do not use water to elute DNA. Incomplete dissociation of DNA from the ChargeSwitch.