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DNA extraction from serum and plasma
for Genetic Research
DNA Extractor SP Kit 1105KB
Wako Catalog No. 296-60501 50 Tests Keep at 2~10℃

Product overview
DNA ExtractorSP Kit is a modified version of DNA Extractor Kit and is designed specifically for extraction of DNA fragments in serum or plasma.
A number of recent studies have reported the presence of DNA fragments in blood originating from cells associated with diseases such as cancer cells.
In Japan and elsewhere, a lot of work is ongoing for development of a genetic diagnosis technique using cell-free DNA in blood for early diagnosis of diseases or monitoring of the patient's condition.
DNA ExtractorSP Kit enables efficient recovery of DNA and is a useful reagent for high-sensitivity detection and analysis of target genes.

Features
  • High DNA recovery rate with the sodium iodide method (almost 100%).
    Efficient recovery with little loss of DNA.
  • Safe procedure.
    Does not require phenol or chloroform treatment.
  • Very low contamination risk.
    The entire procedure can be completed in a single tube.
  • Completely removes lipids from blood samples
    Uses an originally prepared alcohol solution.
  • No variation between samples.

Code No. Product name Volume
296-60501 DNA Extractor SP Kit For 50 uses


  Kit components
  1. Enzyme reaction solution 10 mL x 1
  2. Protease solution 250 μL x 1
  3. Sodium iodide solution 15 mL x 1
  4. Alcohol solution 30 mL x 1
  5. Washing solution (A) 50 mL x 1
  6. Washing solution (B) 50 mL x 1

    Reactions
50 reactions (when the amount of sample is 100 μL)
25 to 30 uses (when the amount of sample is 200 μL)

  Procedure
100 μL (200 μL) *1) of serum or plasma*2)
   ⇓
   ⇓ Add 200 μL (300 μL) of enzyme reaction solution
   ⇓ Add 5 μL (8 to 10 μL) of protease solution*3)
   ⇓ Vortex
   ⇓ Incubate at 56°C for 10 minutes (56°C for 30 minutes)
   ⇓ Add 300 μL (400 μL) of sodium iodide solution
   ⇓ Mix gently
   ⇓ Add 600 μL (900 μL) of alcohol solution
   ⇓ Vortex
   ⇓ at room temperature for 10 minutes
   ⇓ 12,000 to 20,000 at room temperature for 10 minutes
   ⇓
Deposition*4)
   ⇓
   ⇓ Add 1 mL (1.5 mL) of washing solution (A) *5)
   ⇓ Centrifuge*6)
   ⇓ 12,000 to 20,000 at room temperature for 5 minutes
   ⇓
Deposition*4)
   ⇓
   ⇓ Add 1 mL (1.5 mL) of washing solution (B)
   ⇓ Centrifuge*6)
   ⇓ 12,000 to 20,000 at room temperature for 5 minutes
   ⇓
Deposition*4)
   ⇓
   ⇓ Dry at 65°C for 5 minutes
   ⇓ Add 10 to 20 μL of TE (pH: 8.0) or D.W. (10 to 20 μL)
   ⇓ Centrifuge*6)
   ⇓ Dissolve at 65°C for 3 to 5 minutes
   ⇓ Centrifuge*7)
   ⇓
DNA solution: store at 20°C;
*1) The values in parentheses indicate the amounts when the procedure is started with 200 μL of serum or plasma.
*2) Handle the serum (plasma) sample on ice. Add the enzyme reaction solution immediately as DNase in serum (plasma) starts catalyzing the hydrolysis of DNA.
*3) Take out from the kit and place on ice before use.
*4) Discard supernatant and place the tube upside down onto paper towel to remove remaining supernatant.
*5) Add the washing solution (A), and keep it at -20°C for a long term storage.
*6) Mix well until deposits are removed from the vessel wall.
*7) Dissolve until no deposits are visible.

    Experimental data 1
Amplification of exon 5 of p53 in DNA extracted from human serum and plasma
By using DNA Extractor SP Kit, DNA was extracted from 10 human serum samples and one human plasma sample, respectively. The extracted DNA was finally resolved in 20 μL of TE (pH 8.0), and 5 μL of that was subject to amplification of p53-Exon 5 region (308 bp). Commercial available kit basesd on a glass binding method) was used as a comparative method.



    Experimental data 2
Amplification of exon 7 of p53 in DNA extracted from human serum
DNA extracted from human serum samples 2 and 3 (used in the analysis described in "Experimental data 1") as well as DNA from another human serum sample (11) were used to amplify exon 7 of p53 (439 bp).




    References

  • Ishizawa, M., Kobayashi, Y., Miyamura, T. and Matsuura, S. : NucleicAcids Res.,22, 1774 (1994)

  • Sozzi, G., Musso, K., Ratcliffe, C., Goldstraw, P., Pierotti, M.A. and Pastorino, U. :Clin Cancer Res., 5, 2689 (1999)

  • Silva, J. M., Dominguez, G., Garcia, J. M., Gonzalez, R., Villanueva,M. J., Navarro, F., Provencio, M., San, Martin, S., Espana, P. and Bonilla,F.: Cancer Res., 59, 3251 (1999)

  • Shao, Z. M.,Wu, J., Shen, Z. Z. and Nguyen, M. : Clin Cancer Res. 7, 2222 (2001)

  • Chang, H. S., Mizukami, K., Yabuki, A., Hossain, M. A., Rahman, M. M., Uddin, M. M., Arai, T. and Yamato, O. : J. Vet. Diagn. Invest., 22, 708 (2010).

Storage temperature
Keep at -20°C

Related products

Product name Usable samples Catalog No. Package size
P53 Primer Exon 5 Human 312-03511 100 tests
P53 Primer Exon 7 Human 316-03531 100 tests

DNA Extractor® Series available from Wako



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