Blood Genomic DNA Extraction Kit

Details for Product No. ABIN5666465, Supplier: Log in to see
Application
DNA Extraction (DEx)
Options
Supplier
Log in to see
Supplier Product No.
Log in to see
Purpose Blood DNA Extraction Kit is designed for the rapid preparation of 3-20 μg genomic DNA from 100 μl-1 ml whole blood.
Characteristics Blood DNA Extraction Kit is designed for the rapid preparation of 3-20 μg genomic DNA from 100 μL-1 mL whole blood. The extraction process consisits of an initial lysis step and a subsequent proteolytic digestion step using proteinase K with a highly efficient process for binding genomic DNA to the surface of a Spin Column membrane. The high-purity gDNA prepared by this kit usually has an OD260/OD280 between 1.8-2.0 and is suited for a variety of applications, including restriction analysis, sequencing, PCR, Southern blotting etc. No organic extraction or alcohol precipitation, whole procedure could be finished in 20 minutes after lysis.
Components Buffer CL, Buffer PS, Buffer BD, Buffer W1, Buffer W2, Elution Buffer, Proteinase K, Spin Column, Collection Tubes
Application Notes Optimal working dilution should be determined by the investigator.
Protocol This kit is suitable for extracting gDNA from whole blood collected in the presence of anti-coagulants.
1. Sample Treatment
A. Use 200 μL blood sample directly. If the sample volume is less than 200 μL, adjust the sample volume to 200 μL using PS.
B. If the sample volume is more than 200 μL, treat the sample with Buffer CL as following: Add 1~2.5 times volume of Buffer CL, mix thoroughly by inverting spin at 10000 rpm for 1 min,and carefully remove as much of the supernatant as possible. Add 200 μL Buffer PS to the pellet and pipette up and down or vortex to resuspend the cells.
C. If whole blood from amphibian or aves used, the amount of the sample shoule be 5-10 μL, add Buffer PS up to 200 μL and then go to step 2.
2. If RNA-free genomic DNA is required, add 4 μL RNaseA (100 mg/mL) and mix the sample by inverting the tube 2~5 times. Incubate the mixture at room temp.for 2 minutes.(Optional)
3. Add 20 μL of proteinase K, mix thoroughly by vortexing.
4. Add 200 μL Buffer BD, mix thoroughly by vortexing and incubate at 55 °C with occasional inverting the tube for 10 min.(Do not add Proteinase K directly to BD Buffer)
5. Add 200 μL ethanol (96-100 % ) and mix thoroughly by vortexing. (A white precipitate may form on adding ethanol).Briefly spin the tube to remove drops from the inside of the lid.
6. Assemble spin column with one of the provided collection tubes and transfer the mixture (including the white precipitate) into the Spin Column and spin at 12000 rpm for 1 min, discard the flow-through and place the spin column into the collection tube.
7. Add 500 μL Buffer W1 (Ensure ethanol(96-100 %)has been added) into the Spin Column,spin at 12000 rpm for 1 min, discard the flow-through and place the spin column into the collection tube.
8. Add 700 μL Buffer W2 (Ensure ethanol(96-100 %)has been added) into the Spin Column,spin at 12000 rpm for 1 min, discard the flow-through and place the spin column into the collection tube.
9. Repeat step 8 again.
10. Spin at 12000 rpm for 2 min and keep the Spin Column at RT for a few minutes to dry the column thoroughly.
11. Place the Spin Column in a new clean 1.5 mL centrifuge tube, add 50-200 μL pre-warmed Elution Buffer (65C) directly to the center of the membrane,and incubate at RT for 2-5 min (The volume of eluted buffer should be more than 50 μL and distilled water ( pH > 7.0) also can be used to elute gDNA).
12. Spin at 12000 rpm for 2 min to elute DNA.(The yield can be improved by pipetting the elution back onto the column and repeating step 11.)
Restrictions For Research Use only