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Blood Genomic DNA Extraction Kit 

Blood DNA Extraction Kit is designed for the rapid preparation of 3-20 µg genomic DNA from 100 μl-1 ml whole blood.
Cat. No. Size Price Quantity  
TB50005A 50 preps USD 120
TB50005B 200 preps USD 380
Tel: +1 (973) 789-8318Email: [email protected]

Kit Contents

Contents Volume
Buffer CL 60 ml
Buffer PS 12 ml
Buffer BD 27 ml
Buffer W1 21 ml
Buffer W2 16 ml
Elution Buffer 12 ml
Proteinase K 1 ml
Spin Column 50 preps
Collection Tube 50 preps

Storage and Stability

2-8°C 12 months

Description

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.

DNA Yield

Sample Amount DNA Yield
Mammalian blood 100 μl-1 ml 3-20 μg
Bird, fish or amphibian blood 5-10 μl 5-20 μg

Important notes before starting

  1. Buffer BD and Buffer W1 contain strong protein denaturant. Wear gloves, lab coat, etc. when handling these buffers.
  2. Make sure no precipitates formed in Buffer BD and Buffer W1. If necessary, warm the solutions and mix well until the solutions become clear again.
  3. All centrifugation steps should be carried out at room temperature (RT, 15-25°C).
  4. RNase A (not provided) can be used if RNA-free DNA is required.
  5. Avoid freeze/thaw cycles of blood sample. Freeze/thaw cycle of blood sample would result in low yield of genomic DNA or DNA length less than 30kb.
  6. Add 7 ml and 64 ml ethanol (96-100%) into Buffer W1 and Buffer W2 respectively when first use.
  7. For research use only. Not intended for human or diagnostic use.

Procedure

Make sure ethanol (96-100%) has been added into Buffer W1 and Buffer W2.
This kit is suitable for extracting gDNA from whole blood collected in the presence of anti-coagulants.

  1. Sample Treatment
    1. Use 200 μl blood sample directly. If the sample volume is less than 200 μl, adjust the sample volume to 200 μl using PS.
    2. 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.
    3. 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.)

    Troubleshooting

    The yield of genomic DNA is low

    This may be caused by incomplete lysis of cells or the sample is old/overgrown.
    1. Incomplete lysis of cells. Try to reduce the amount of samples and too many cells may results in incomplete lysis of cells and also clogging the spin column.
    2. The sample is old. It may be necessary to use fresh samples. If users need to process the preparation the other day; pellet the cells, remove the culture medium and store the pellet at -80°C.
    3. Make sure ethanol (96-100%) had been added into Buffer W1 and Buffer W2.

    Downstream application failure with gDNA eluate

    Keep the Spin Column at room temperature for a few minutes to thoroughly dry the column before adding elution buffer and the elution buffer should be added directly to the center of the membrane.

Datasheet Download

Blood DNA Extraction Kit

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