GenepHlow™ Gel/PCR Kit (DFH100, DFH300)

Gel Extraction Kit | PCR Cleanup Kit
GenepHlow™ Gel/PCR Kit (DFH100, DFH300)
The GenepHlow™ Gel Extraction Kit and PCR Cleanup Kit was designed to recover or concentrate DNA fragments from agarose gel, PCR or other enzymatic reactions. This gel extraction kit and PCR cleanup kit includes a pH indicator which is premixed with the binding buffer to ensure optimal pH, facilitate DNA binding and allow for easy observation of undissolved agarose gel. If pH exceeds the optimal level (>7.5), the color of the solution will appear purple instead of yellow. 3M Sodium Acetate (pH5.0), which is included with the kit, can then be added to the solution to adjust pH and return the color to yellow. Chaotropic salt is used to dissolve agarose gel and denature enzymes. DNA fragments in the chaotropic salt are bound by the glass fiber matrix of the spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified DNA fragments are eluted by a low salt Elution Buffer, TE or water. The pH indicator, salts, enzymes and unincorporated nucleotides can be effectively removed from the reaction mixture without phenol extraction or alcohol precipitation and the purified DNA is ready for use in subsequent reactions.

 

 

Advantages (Cat. # DFH100, DFH300)
  • High Efficiency: up to 90% recovery from agarose gels, up to 95% recovery from PCR or other enzymatic reactions
  • Only 20 minute gel extraction and 10 minute PCR cleanup
  • Convenient: Gel/PCR binding buffer is premixed with a pH indicator to allow for easy determination of optimal pH
  • Includes 3M Sodium Acetate (pH5.0) to adjust pH if it becomes too high
  • Sample Size: up to 300 mg of agarose gel slice or  up to 100 μl of PCR products
  • Broad Fragment Size Range: 70 bp–20 kb
  • Spin Columns: glass fiber membrane optimized for both gel extraction and PCR cleanup
  • Elution Volume: 20-50 µl
  • Storage: dry at room temperature (15-25°C)

Applications

Fluorescent and Radioactive Sequencing, PCR, Restriction Enzyme Digestion, DNA Labeling and Ligation, Microarray, Transfection

Components

  • Gel/PCR Buffer (premixed with a pH indicator)
  • 3M Sodium Acetate (pH5.0)
  • W1 Buffer
  • Wash Bufffer
  • Elution Buffer (10 mM Tris-HCl, pH8.5 at 25ºC)
  • DFH Columns
  • 2 ml Collection Tubes

Quality Control

The quality of the GenepHlow™ Gel/PCR Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. DNA fragments are purified from either agarose gel, PCR products or other aqueous solutions analyzed by electrophoresis.

GenepHlow™ Gel/PCR Kit Interactive Protocol

1. Gel Extraction

2. PCR Cleanup

GenepHlow™ Gel/PCR Kit Functional Test Data 

Figure 1. Gel slice DNA fragments ranging from 250 bp-10 kb were extracted using the GenepHlow™ Gel/PCR Kit (lane 1, 2, 3). The purified DNA from a 50 µl eluate was analyzed by electrophoresis on a 1% agarose gel.
M = Geneaid 1 Kb DNA Ladder (control, total DNA = 1100 ng)
Test DNA Conc. 260/280 Yield Recovery
1 18.2 ng/µl 1.84 910 ng 82%
2 18.7 ng/µl 1.82 935 ng 85%
3 19.2 ng/µl 1.82 960 ng
87.3%
Figure 2. PCR product DNA fragments ranging from 250 bp-10 kb were extracted using the GenepHlow™ Gel/PCR Kit (lane 1, 2, 3). The purified DNA from a 50 µl eluate was confirmed by spectrophotometer and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder (control, total DNA = 1073 ng)
Test DNA Conc. 260/280 Yield Recovery
1 19.7 ng/µl 1.86 985 ng 91.8%
2 20.3 ng/µl 1.85 1015 ng 94.6%
3 20.5 ng/µl 1.86 1025 ng 95.5%
Optimal pH  
pH Too High
Figure 3. A pH indicator is premixed with the binding buffer to ensure optimal pH, facilitate DNA binding and allow for easy observation of undissolved agarose gel. If pH exceeds the optimal level (>7.5), the color of the solution will appear purple instead of yellow. 3M Sodium Acetate (pH5.0), which is included with the kit, can then be added to the solution to adjust pH and return the color to yellow.

Gel Extraction and PCR Cleanup Kits Specifications

Specifications
 
Cat. #
DFG100
DFG300
DFC100
DFC300
DFH100
DFH300
DFM010
DFM025
96DFG04
96DFG10
DF101
DF301
DFL100
DFL300
Format
Spin Column
Spin Column
Spin Column
Maxi Spin Column
96 Well Plate
Spin Column
Silica Matrix
Binding Capacity 10 µg 10 µg 10 µg 200 µg 10 µg 10 µg 5 µg
Sample Agarose Gel Slice PCR Product Agarose Gel Slice/PCR Product Agarose Gel Slice/Restriction Enzyme Digest Product Agarose Gel Slice Agarose Gel Slice/PCR Product Agarose Gel Slice/PCR Product
Fragment Size 70 bp-20 kb 70 bp-20 kb 70 bp-20 kb 100 bp-10 kb 70 bp-20 kb 40-200 bp 100 bp-50 kb
pH Indicator Yes Yes Yes Yes N/A N/A Yes
Recovery Up to 90% Up to 95% Up to 95% Up to 80% Up to 80% Up to 95%
Up to 95% (<10 kb)
Up to 80% (10-50 kb)
Elution Volume 20-50 µl 20-50 µl 20-50 µl 500-1000 µl 60-80 µl 20-50 µl 10-20 µl
Operation Time 20 minutes 10 minutes <20 minutes <25 minutes 45 minutes <20 minutes <20 minutes

Publications

 

 
 

 

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