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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.

Quick Protocol

 

 

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)
TestDNA Conc.260/280YieldRecovery
118.2 ng/µl1.84910 ng82%
218.7 ng/µl1.82935 ng85%
319.2 ng/µl1.82960 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.260/280YieldRecovery
119.7 ng/µl1.86985 ng91.8%
220.3 ng/µl1.851015 ng94.6%
320.5 ng/µl1.861025 ng95.5%
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.

PCR Cleanup Kits Specifications

Specifications
 
Presto™ 96 Well PCR Cleanup Kit 
Cat. #
DFC100
DFC300
DFH100
DFH300
DFM010
DFM025
DFI100
DFI300
DF101
DF301
DFL100
DFL300
96DFH04
96DFH10
Format
Spin Column
Spin Column
Maxi Spin Column
Midi Spin Column
Spin Column
Silica Matrix96 Well Plate
Binding Capacity10 µg10 µg200 µg40 µg10 µg5 µg10 µg
SamplePCR ProductAgarose Gel Slice/PCR ProductAgarose Gel Slice/Restriction Enzyme Digest ProductPCR ProductAgarose Gel Slice/PCR ProductAgarose Gel Slice/PCR ProductPCR Product
Fragment Size70 bp-20 kb70 bp-20 kb100 bp-10 kb100 bp-20 kb40-200 bp100 bp-50 kb70 bp-20 kb
pH IndicatorYesYesYesYesN/AYesN/A
RecoveryUp to 95%Up to 95%Up to 80%Up to 90%Up to 95%
Up to 95% (<10 kb)
Up to 80% (10-50 kb)
Up to 95%
Elution Volume20-50 µl20-50 µl500-1000 µl30-50 µl20-50 µl10-20 µl60-80 µl
Operation Time10 minutes<20 minutes<25 minutes10 minutes<20 minutes<20 minutes20 minutes

Related Gel Extraction and PCR Cleanup Products

Product Name
RXNS
Catalogue Number
100, 300
DFG100/300
100, 300
DFC100/300
100, 300
DFH100/300
10, 25
DFM010/25
4 x 96, 10 x 96
96DFH04/10
100, 300
DF101/301
100, 300
DFL100/300
100, 300
DF100/300
Presto™ 96 Well Gel Extraction Kit4 x 96, 10 x 9696DFG04/10
GenepHlow™ DNA Cleanup Midi Kit100, 300DFI100/300