DNA Cleanup Maxi Kit (DFM010, DFM025)

PCR Cleanup Maxi Kit | Gel Extraction Kit Maxi
DNA Cleanup Maxi Kit (DFM010, DFM025)
GenepHlow™ DNA Cleanup Maxi Kits were designed to recover or concentrate DNA fragments from large volume agarose gel, restriction enzyme digestion or PCR reaction products. Gel/PCR Buffer (yellow color indicating optimal pH≤7.5) is premixed with a pH indicator 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 while DNA fragments 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. # DFM010, DFM025)
  • High Efficiency: up to 80% recovery
  • 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 5 g of agarose gel slice or up to 2 ml of restriction enzyme digestion product or PCR product
  • Broad Fragment Size Range: 100 bp–10 kb
  • Operation Time: 24 minutes for enzymatic reaction cleanup and 36 minutes for gel extraction
  • Spin Columns: glass fiber membrane
  • Elution Volume: 500-1,000 µl (50 µl dead volume)
  • Binding Capacity: 200 µg
  • Storage: dry at room temperature (15-25ºC) 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

  • 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)
  • DM Columns

Quality Control

The quality of the GenepHlow™ DNA Cleanup Maxi 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, restriction enzyme digestion products, PCR products or other aqueous solutions analyzed by electrophoresis.

GenepHlow™ DNA Cleanup Maxi Kit Functional Test Data 

Figure 1. 200 µg of pbluescript plasmid DNA was digested by Hind III endonuclease at 37ºC for 2 hours. Following digestion, the linear DNA fragment was purified using the GenepHlow™ DNA Cleanup Maxi Kit. The unpurified DNA fragment (lane 1) and 3 replications of 5 µl purified linear DNA fragments (lanes 2-4) were loaded on 0.8% agarose gel. M = Geneaid 1 Kb DNA ladder
Sample ng/µl 260/280 260/230 Elution Yield (µg) Recovery
2 - Purified DNA 155.4 ng/µl 1.87 2.17 1000 µl 155.4 77.7%
3 - Purified DNA 157.8 ng/µl 1.86 2.22 1000 µl 157.8 78.9%
4 - Purified DNA 152.6 ng/µl 1.87 2.24
1000 µl
152.6 76.3%
Optimal pH  
pH Too High
Figure 2. 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

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