PCR Cleanup Kit (DFC100/DFC300)

PCR Cleanup Kit
PCR Cleanup Kit (DFC100/DFC300)

The GenepHlow™ PCR Cleanup Kit was designed to recover, concentrate or purify DNA fragments from PCR or other enzymatic reactions. This PCR purification kit includes a pH indicator as an optional addition to the PB binding buffer to ensure optimal pH and facilitate DNA binding. 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 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. The GenepHlow™ PCR Cleanup Kit is a cost effective, high performance alternative for efficient DNA fragments PCR purification.

 

 

Advantages (Cat. # DFC100, DFC300)
  • High Efficiency: up to 95% recovery from PCR or other enzymatic reactions
  • Only 10 minute PCR cleanup
  • Includes a pH Indicator for easy determination of optimal pH
  • Includes 3M Sodium Acetate (pH5.0) to adjust pH if it becomes too high
  • Sample Size: up to 100 μl of PCR products
  • Broad Fragment Size Range: 70 bp–20 kb
  • Spin Columns: glass fiber membrane optimized for PCR cleanup
  • Elution Volume: 20-50 µl
  • Storage: dry at room temperature (15-25°C) for up to 18 months without showing any reduction in performance

Applications

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

Components

  • PB Buffer
  • ph Indicator
  • 3M Sodium Acetate (pH5.0)
  • 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™ PCR Cleanup Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. DNA fragments are purified from PCR products and analyzed by electrophoresis.

GenepHlow™ PCR Cleanup Kit Functional Test Data                

Figure 1. PCR product DNA fragments ranging from 250 bp-10 kb were extracted using the GenepHlow™ PCR Cleanup 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 2. A pH indicator is included as an optional additon to the PB binding buffer to ensure optimal pH and facilitate DNA binding. 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 Matrix 96 Well Plate
Binding Capacity 10 µg 10 µg 200 µg 40 µg 10 µg 5 µg 10 µg
Sample PCR Product Agarose Gel Slice/PCR Product Agarose Gel Slice/Restriction Enzyme Digest Product PCR Product Agarose Gel Slice/PCR Product Agarose Gel Slice/PCR Product PCR Product
Fragment Size 70 bp-20 kb 70 bp-20 kb 100 bp-10 kb 100 bp-20 kb 40-200 bp 100 bp-50 kb 70 bp-20 kb
pH Indicator Yes Yes Yes Yes N/A Yes N/A
Recovery Up 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 Volume 20-50 µl 20-50 µl 500-1000 µl 30-50 µl 20-50 µl 10-20 µl 60-80 µl
Operation Time 10 minutes <20 minutes <25 minutes 10 minutes <20 minutes <20 minutes 20 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 Kit 4 x 96, 10 x 96 96DFG04/10
GenepHlow™ DNA Cleanup Midi Kit 100, 300 DFI100/300