DNA Cleanup Midi Kit (DFI100, DFI300)

DNA Cleanup Midi Kit
DNA Cleanup Midi Kit (DFI100, DFI300)
GenepHlow™ DNA Cleanup Midi Kits were designed to recover or concentrate DNA fragments from PCR or other enzymatic reactions. This DNA cleanup 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 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. # DFI100, DFI300)
  • High Efficiency: up to 90% recovery
  • Convenient: includes pH indicator for easy determination of optimal pH and sodium acetate to adjust pH if it becomes too high
  • Includes 3M Sodium Acetate (pH5.0) to adjust pH if it becomes too high
  • Sample Size: 10-40 µg of DNA in up to 200 µl PCR/Digestion solutions
  • Broad Fragment Size Range: 100 bp–20 kb
  • Operation Time: 10 minutes
  • Spin Columns: glass fiber membrane
  • Elution Volume: 30-50 µl
  • Storage: dry at room temperature (15-25°C) for up to 18 months without showing any reduction in peformance

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)
  • DF Midi Columns
  • 2 ml Collection Tubes

Quality Control

The quality of the GenepHlow™ DNA Cleanup Midi Kit is tested on a lot-to-lot basis by purifying DNA fragments of various sizes from PCR products, digestion products or other aqeous solutions. The purified DNA is analyzed by electrophoresis.
 
GenepHlow™ DNA Cleanup Midi Kit Functional Test Data 
Figure 1. 40 µg of pbluescript plasmid DNA was digested by Hind III endonuclease at 37ºC for 1 hour. Following digestion, the linear DNA fragment was purified using the GenepHlow™ DNA Cleanup Midi 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 731.4 ng/µl 1.84 2.28 50 µl 36.6 92%
3 - Purified DNA 724.8 ng/µl 1.83 2.24 50 µl 36.2 91%
4 - Purified DNA 730.5 ng/µl 1.84 2.23
50 µl
36.5 91%
Optimal pH  
pH Too High
Figure 2. A pH indicator is included 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.

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
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DFC100/300
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DFH100/300
10, 25
DFM010/25
4 x 96, 10 x 96
96DFH04/10
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DF101/301
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DFL100/300
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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