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GenepHlow™ Gel/PCR Kit (DFH100, DFH300)

DNA Extraction/Gel Extraction

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.

DOCUMENTS

DESCRIPTION

Specifications (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
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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%
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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%

 

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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

 
SpecificationsGenepHlow™ Gel Extraction KitGenepHlow™ PCR Cleanup KitGenepHlow™ Gel/PCR KitGenepHlow™ DNA Cleanup Maxi KitPresto™ 96 Well Gel Extraction KitSmall DNA Fragments Extraction KitLarge DNA Fragments Extraction Kit
Cat. #DFG100
DFG300
DFC100
DFC300
DFH100
DFH300
DFM010
DFM025
96DFG04
96DFG10
DF101
DF301
DFL100
DFL300
FormatSpin ColumnSpin ColumnSpin ColumnMaxi Spin Column96 Well PlateSpin ColumnSilica Matrix
Binding Capacity10 µg10 µg10 µg200 µg10 µg10 µg5 µg
SampleAgarose Gel SlicePCR ProductAgarose Gel Slice/PCR ProductAgarose Gel Slice/Restriction Enzyme Digest ProductAgarose Gel SliceAgarose Gel Slice/PCR ProductAgarose Gel Slice/PCR Product
Fragment Size70 bp-20 kb70 bp-20 kb70 bp-20 kb100 bp-10 kb70 bp-20 kb40-200 bp100 bp-50 kb
pH IndicatorYesYesYesYesN/AN/AYes
RecoveryUp 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 Volume20-50 µl20-50 µl20-50 µl500-1000 µl60-80 µl20-50 µl10-20 µl
Operation Time20 minutes10 minutes<20 minutes<25 minutes45 minutes<20 minutes<20 minutes

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