It has been added to the cart.

It has been added to the comparison table.

DNA ExtractorR Kit

for Genetic Research
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
  • Structural Formula
  • Label
  • Packing
SDS
Comparison
Product Number
Package Size
Price
Inventory
Distributor
295-50201
Barcode No
4987481359765
50Tests
Inquire

In stock

Document

SDS ...all
Product Specification Sheet
Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Kit component

50 tests

Sodium Iodide Solution 1 x 26 mL
Sodium N-Lauryl Sarcosinate Solution 1 x 1.2 mL
Washing Solution (A) 1 x 42 mL
Washing Solution (B) 2 x 40 mL
Glycogen Solution 1 x 0.1 mL

Outline

Residual DNA testing for vaccines and biopharmaceuticals

This is sodium iodide method based DNA extraction kit to isolate residual host cell DNA from biological samples. This kit follows the residual DNA extraction protocols described in the United States Pharmacopeia (USP) 43-NF38, <509> Residual DNA Testing.
The isolated DNA can be used for qPCR. This is suitable for the residual DNA testing derived from host cells including CHO cells, E.coli and yeast.
This kit can also be used for DNA isolation for Threshold® System provided by Molecular Devices, LLC, which can quantify total DNA regardless of the origin.

Virus DNA quantification in human serum

This can be used for virus DNA isolation from human serum.

Features

  • Efficient extraction of trace DNA (100-1,000 fg)
  • Perform in a single tube for whole process
  • Extraction complete in 60-90 minutes
  • Suitable for quantification by qPCR and Threshold® System (Molecular Devices, LLC)
  • Pretreatment protocol for samples containing high concentration protein

Basic operation of DNA extraction by sodium iodide method

  1. Add sodium iodide, a chaotropic ion and detergents to solubilize protein and lipids.
  2. Add 2-propanol and DNA is coprecipitated with glycogen.
  3. Collect DNA pellets.

Application

Spike and recovery test of CHO cell-derived DNA

Examined the recovery rate of CHO cell-derived DNA from spiked culture supernatant to evaluate the extraction efficiency.

Methods
  1. Added 10 fg to 1 ng of CHO cell-derived DNA to culture supernatant of PANC-1 cells.
  2. Extracted DNA from the supernatant with this kit following the manual.
    -Protocol: Protocol #2
    -Pretreatment: None
  3. Conducted qPCR and measured the Cq value of the extracted DNA. The same measurement was conducted for purified water spiked with CHO cell-derived DNA without DNA extraction, this was used as standard condition.
  4. Create a standard curve using the mean Cq values of the standard condition, and calculated the recovery rate.
Samples
  1. Purified water containing CHO cell-derived DNA (no DNA extraction): Standard conditions
  2. DNA extracted from the culture supernatants of PANC-1 cells containing CHO cell-derived DNA by this kit: Culture supernatants
qPCR reagents
  • GeneAce SYBR® qPCR Mix α No ROX (Nippon Gene#319-07703)
  • Optical Flat 8–Cap Strips for 0.2 mL tube strips/Plates (BIO-RAD#TCS0803)
  • Hard-Shell® Thin-wall 96-well PCR plates (BIO-RAD#HSP9601)
Results
1. Standard conditions 2. Culture supernatants
Spike DNA (fg) Cq 1 Cq 2 Mean Cq 1 Cq 2 Mean
0 ND
10
100 36.89 ND 36.89 37.44 36.73 37.09
1,000 33.44 33.91 33.68 33.94 34.09 34.01
10,000 30.23 30.17 30.20 30.72 30.96 30.84
100,000 26.74 26.68 26.71 26.90 27.03 26.96
1,000,000 23.39 23.28 23.33 23.61 23.48 23.54

ND: Not detected

00071984_img01R.png
Good extraction efficiency of CHO-derived DNA from culture supernatant within the range of 100 fg to 1 ng.

Spike and recovery test of E. coli-derived DNA

Examined the recovery rate of E. coli-derived DNA from spiked sample to evaluate the extraction efficiency.

Methods
  1. Added 10 fg to 1 ng of E.coli-derived DNA to purified water.
  2. Extracted DNA from the spiked sample with this kit following the manual.
    -Protocol: Protocol #1
  3. Conducted qPCR and measured the Cq value of the extracted DNA. The same measurement was conducted for the spiked sample before DNA extraction.
  4. Create a standard curve using the mean Cq values of the spiked samples before extraction, and calculated the recovery rate after the extraction.
Samples

Purified water spiked with E. coli-derived DNA

qPCR reagents
  • GeneAce SYBR® qPCR Mix α No ROX (Nippon Gene#319-07703)
  • COptical Flat 8–Cap Strips for 0.2 mL tube strips/Plates (BIO-RAD#TCS0803)
  • CHard-Shell® thin-wall 96-well PCR plates (BIO-RAD#HSP9601)
Results
00071984_img02.png
Good extraction efficiency of E.Coli-derived DNA within the range of 1 pg to 1 ng.

Spike and recovery test from Human serum sample

λ/HindⅢ 10 to 1,000 pg was labeled with 32P and added to 100 μL of human serum. DNA was extracted using this kit and the recovery rate was calculated.

00071984_img03.png

Reference

Ishizawa, M. et al., Nucleic Acids Res., 19, 5792 (1991).

Good extraction efficiency from serum sample.

Detection of virus DNA from human serum by PCR

Serum containing hepatitis B virus (HBV) was diluted in stages. DNA was extracted using this kit and the phenol method. HBV-DNA was amplified with PCR, and DNA was detected using agarose gel electrophoresis.

00071984_img04.png

Lane 1: Marker DNA(φⅩ174 DNA/HaeⅢ)

HBV-DNA was detected with higher specificity and sensitivity using this kit than phenol method.

Evaluation For Antibody Drug

DNA extraction from high concentration protein-containing solution

Conducted DNA spike and recovery test from various concentration of IgG solution.

Methods
  1. Added CHO cell-derived DNA (final DNA concentration: 2 pg/mL) to 10 – 100 mg/mL of IgG solution.
  2. Extracted DNA from the spiked IgG solution with this kit following the manual.
    -Protocol: Protocol #2
    -Pretreatment: Protocol for the sample containing more than 2 mg/mL of protein (Proteinase K treatment)
  3. Measured the recovery rate by qPCR
Results
Recovery rate of CHO cell-derived DNA from IgG solution
00071984_img05.png
Cq value of IgG solution at various concentration
Sample Input Protein Concentration (mg/mL)
10 20 40 60 80 100
Cq 31.596 31.566 31.351 31.726 31.502 31.282 31.658
31.599 31.608 31.774 31.787 31.485 31.660
31.338 31.861 31.500 31.850 31.560 31.369
Average Cq 31.511 31.678 31.542 31.788 31.531 31.379 31.659
Difference of Cq 0 -0.167 0.136 -0.246 0.257 0.152 -0.280
Power 1 0.891 1.099 0.843 1.195 1.111 0.824
Recovery Rate (%) 100 89.1 109.9 84.3 119.5 111.1 82.4
Good DNA extraction efficiency from solution containing high concentration protein (100 mg/mL).

DNA extraction from antibody drug-mimic solution

Conducted DNA spike and recovery test from IgG solutions made of various types of buffers generally used in antibody drug and from IgG solutions containing various additives used in antibody drug.

Methods
  1. Made antibody drug-mimic solutions by adding 20 mg/mL of IgG to various buffers generally used in antibody drug (Table 1. ①-⑥) and to phosphate buffer containing various additives (Table 2. ①-⑩). Then add CHO cell-derived DNA (final DNA concentration: 20 pg/mL) to those antibody drug-mimic solutions.
  2. Extracted DNA from the spiked samples with this kit following the manual.
    -Protocol: Protocol #2
    -Pretreatment: Protocol for the sample containing more than 2 mg/mL of protein (Proteinase K treatment)
  3. Measured the recovery rate by qPCR.
  • Table 1. Buffer Composition
    No. Buffer Composition
    10mM PB pH6.0, 0.15M NaCl, 2mM EDTA
    10mM PB pH7.4, 0.15M NaCl, 2mM EDTA
    50mM NaOAc pH5.2, 0.15M NaCl, 2mM EDTA
    10mM PB pH6.0, 2mM EDTA
    10mM PB pH7.4, 2mM EDTA
    50mM NaOAc pH5.2, 2mM EDTA
  • Table 2. Additive-containing Buffer

    Buffer: 10mM PB pH6.0, 0.15M NaCl, 2mM EDTA

    No. Additive & Concentration
    Sucrose (10w/v%)
    Trehalose Dihydrate (10w/v%)
    D(-)-Mannitol (10w/v%)
    D(-)-Sorbitol (10w/v%)
    D(+)-Maltose Monohydrate (10w/v%)
    L(+)-Arginine Hydrochloride (1w/v%)
    L‐Histidine (1w/v%)
    Glycine (1w/v%)
    Polyoxyethylene(20) Sorbitan Monooleate (Tween80) (0.5v/v%)
    Polyoxyethylene(20) Sorbitan Monolaurate (Tween20) (0.5v/v%)
Results
DNA Recovery Rate from each buffer
00071984_img06.png
Cq values of each buffer
Sample Input Buffer
1 2 3 4 5 6
Cq 31.473 31.693 31.544 31.218 31.058 31.595 31.181
31.364 31.196 30.973 31.568 31.085 31.343 31.112
31.045 31.530 31.787 31.315 30.942 31.452 31.558
Average Cq 31.294 31.473 31.435 31.367 31.028 31.463 31.284
Difference of Cq 0 -0.179 -0.141 -0.073 0.266 -0.169 0.010
Power 1 0.883 0.907 0.951 1.203 0.890 1.007
Recovery Rate (%) 100 88.3 90.7 95.1 120.3 89.0 100.7
Cq values of each additive-containing buffer
Sample Input Additive-containing buffer
Cq 31.473 31.826 31.327 30.907 31.143 31.071 31.525 31.525 31.013 31.086
31.364 31.194 30.991 31.759 31.103 31.279 31.068 31.442 31.203 31.340 30.870
31.045 30.703 31.009 30.978 31.225 31.479 30.757 31.020 31.020
Average Cq 31.294 31.241 31.159 31.225 31.123 31.109 31.273 31.461 31.162 31.124 30.992
Difference of Cq 0 0.053 0.135 0.069 0.171 0.185 0.021 -0.167 0.132 0.170 0.302
Power 1 1.037 1.098 1.049 1.126 1.137 1.015 0.891 1.096 1.125 1.233
Recovery Rate (%) 100 103.7 109.8 104.9 112.6 113.7 101.5 89.1 109.6 112.5 123.3
DNA Recovery Rate from each additive-containing buffer
00071984_img07.png
Good DNA extraction efficiency from various buffer and additive.

Overview / Applications

Outline This is for research use only. Do not administer it to human. Molecular Biology-DNA Extraction Kits. Detects and quantitates contaminant DNA in serum and residual DNA in biopharmaceuticals. Employs a new extraction procedure for DNA purification from a single tube. This procedure, using Sodium Iodide (NaI) as a chaotropic agent, realizes DNA isolation of both high quality and high recovery from biological fluids without the use of phenol or chloroform. Complex and laborious manipulations are avoided when using this kit. In addition, this kit has a modified application for use with Molecular Devices' Threshold(TM) System.
PRINCIPLE of DNA EXTRACTION: A high concentration of chaotropic reagent, NaI, and an anionic detergent participate in solubilization of the proteins and lipids contained in biological samples. After addition of isopropanol to the mixture, nucleic acids are co-precipitated with polysaccharide glycogen as a carrier, while other components remain soluble in the solution phase.

Ref.: Ishizawa, M., Kobayashi, T. and Matsuura, S., ''Simple procedure of DNA Isolation from Human Serum'', Nucleic Acids Res., 19, 5792 (1991).
Hui Cai, Xuelin Gu, Mary S. Scanlan, and ChrisR. Lively,: Development of a quantitative PCR assay for residual mouse DNA and comparison of four sample purification methods for DNA isolation., Journal of Pharmaceutical and Biomedical Analysis, 55, 71-77 (2011).
This method was listed on USP39-NF34, page 1413/Pharmacopeial Forum. Vol.41(4).
Purpose DNA extraction.

Property

Manufacturer Information

Alias

For research use or further manufacturing use only. Not for use in diagnostic procedures.

Product content may differ from the actual image due to minor specification changes etc.

If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.

+49-2131-311-271

Reception hours: Mon-Fri 9:00 - 15:00 (CET)For other hours than the above, please contact us via the inquiry form.