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Description : The EZgeneTM Forensic gDNA Purification Kit provides a rapid and easy method for the isolation of genomic DNA from forensic samples, such as dry blood, buccal swabs, and sperm for consistent PCR and Southern analysis. This kit can also be used for the preparation of genomic DNA from mouse tail snips, whole blood, buffy coat, serum, and plasma. No phenol/chloroform extractions and isopropanol or ethanol precipitation are needed. DNA purified using this method is ready for downstream applications such as PCR, Southern blotting, and restriction digestion. In this procedure, samples are first lysed and applied to the spin column that DNA binds. While cellular debris, hemoglobin, and other proteins are effectively washed away by DNA Wash Buffer, pure DNA is eluted in sterile deionized water or elution buffer. Each ezBind column can bind approximately 100 μg DNA. Once reconstituted, Protease K must be stored at -20°C. All other components can be stored at 22-25°C.Under these conditions, performance of all components of the kit are guaranteed at least 12 months from the date of purchase. Under cool ambient conditions, a precipitate may form in the Buffer BL; heat the bottle at 37°C to dissolve the precipitate before use. Buffer BL contains acidic acid and chaotropic salts, which may form reactive compounds when combines with bleach. Do not add bleach or acidic solutions directly to the preparation waste. Ware gloves and protective eyeware when handling this solution. Product GD2512-00 GD2512-01 GD2512-02 Preps 4 50 250 DNA Columns 4 50 250 2 mL Collection Tubes 8 100 500 Buffer BL 3 mL 30 mL 150 mL Buffer TL 3 mL 20 mL 100 mL Buffer KB 3 mL 28 mL 135 mL DNA Wash Buffer 2 mL 15 mL 3 x 24 mL Elution Buffer 2 mL 15 mL 60 mL Protease K 3 mg 30 mg 5 x 30 mg User Manual 1 1 1 For isolation gDNA from sperm Product GD2512-00 GD2512-01 GD2512-02 10x Buffer A 5 mL 50 mL 250 mL Buffer B 1 mL 15 mL 70 mL Note: The kit is supplied with enough buffers for the standard protocol. However, due to increased volumes called for in some protocols (such as the buccal swab protocol), extra buffers can be purchased separately from Biomiga. See product inforamtion on our website or call customer service for price information. The total DNA yield can be determined by, [DNA] = (Absorbance260) x (0.05 μg/ μL) x (Dilution factor) The quality of DNA can be evaluated by OD260/280. A ratio of (A260/A280) of 1.8-1.9 corresponds to 90%-95% purity. Expected yields vary with both amount, and type of tissue used. 30 mg of fresh tissue will yield approximately10-40 μg DNA with two elutions (each 100 μL). Problem Possible Cause Suggestions Colored residue in column After washing Forgot to add ethanol Before applying sample to column, both Buffer BL and ethanol must be added. See protocol above. Forgot to add ethanol to DAN Wash Buffer Dilute DNA Wash Buffer with the indicated volume of absolute ethanol before use. Incomplete lysis due to improper mixing with Buffer BL. Buffer BL is viscous and the sample must be vortexed thoroughly. No ethanol added to DNA Wash Buffer Concentrate. Dilute DNA Wash Buffer with the indicated volume of absolute ethanol before use. Column clogged Incomplete lysis Extend incubation time of lysis with Buffer TL and protease. Add the correct volume of Buffer BL and incubate for specified time at 70℃. It may be necessary to extend incubation time by 10 min. Sample too large If using more than 30 mg tissue, increase volumes of Proteinase K, Buffer TL, Buffer BL, and ethanol. Pass aliquots of lysate through one column successively. Sample too viscous Divide sample into multiple tubes, adjust volume to 250 μL with 10 mM Tris-HCl. Low DNA yield Clogged column See above Poor sample release from collection paper Incubate the OB specimen collection paper longer in TL buffer. Shake the tubes frequently. Poor elution Repeat elution or increase elution volume. Incubation of column at 70℃ for 5 min with Elution Buffer may increase yields. Improper washing DNA Wash Buffer Concentrate must be diluted with absolute (100%) ethanol as specified on Page 3 before use. Trouble Shooting Guide (Continue ) Problems Possible Cause Suggestions A260 /A280 ratio lower than 1.7 Extended centrifugation during elution step. Resin from the column may be present in eluate. Avoid centrifugation at speeds higher than specified. The material can be removed from the eluate by centrifugation - it will not interfere with PCR or restriction digests. Poor cell lysis due to incomplete mixing with Buffer BL Repeat the procedure, this time making sere to vortex the sample with Buffer BL immediately and completely. Incomplete cell lysis or protein degradation due to insufficient incubation. Increase incubation time with Buffer TL and protease. Ensure that no visible pieces of tissue remain. Samples are rich in protein. After applying to column, wash with 300 μL of a 1:1 mixture of Buffer BL and ethanol and then with DNA Wash Buffer. No DNA eluted Poor cell lysis due to improper mixing with Buffer BL. Mix thoroughly with Buffer BL prior to loading ezBind column. Catalog # Product Name Preps Price $ GD2211-01 Tissue gDNA kit 50 90.00 GD2211-02 Tissue gDNA kit 250 420.00 GD2814-01 96-well tissue gDNA kit 4x96 680.00 GD2814-02 96-well tissue gDNA kit 20x96 3000.00 GD2311-01 Blood gDNA mini kit 50 90.00 GD2311-02 Blood gDNA mini kit 250 420.00 GD2312-01 Blood gDNA midi kit 10 90.00 GD2312-02 Blood gDNA midi kit 25 200.00 GD2314-01 Blood gDNA maxi kit 10 160.00 GD2314-02 Blood gDNA maxi kit 25 360.00 GD2815-01 96-well blood gDNA kit 4x96 680.00 GD2815-02 96-well blood gDNA kit 20x96 3000.00 GD2411-01 Bacterial gDNA kit 50 110.00 GD2411-02 Bacterial gDNA kit 250 495.00 GD2412-01 Soil gDNA kit 50 90.00 GD2412-02 Soil gDNA kit 250 420.00 GD2413-01 Insect gDNA kit 50 90.00 GD2413-02 Insect gDNA kit 250 420.00 GD2414-01 Mullusc gDNA kit 50 90.00 GD2414-02 Mullusc gDNA kit 250 420.00 GD2415-01 Yeast gDNA kit 50 90.00 GD2415-02 Yeast gDNA kit 250 420.00 GD2416-01 Fungal gDNA kit 50 90.00 GD2416-02 Fungal gDNA kit 250 420.00 GD2512-01 Forensic gDNA purification kit 50 85.00 GD2512-02 Forensic gDNA purification kit 250 380.00Introduction
Storage and Stability
Safety Information
Kit Contents
Before Starting(Important):
Determination of Yield and Quality
Trouble Shooting Guide
Related EZgeneTM Products
- Model: GD2512-02
- Manufactured by: Biomiga
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