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DNA Cleanup Midi Kit (DFI100, DFI300)

DNA Extraction/PCR Cleanup

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.

DOCUMENTS

DESCRIPTION

Specifications (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)

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

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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
Purified DNA (Lane-2)
731.4 ng/µl
1.84
2.28
50 µl
36.6
92%
Purified DNA (Lane-3)
724.8 ng/µl
1.83
2.24
50 µl
36.2
91%
Purified DNA (Lane-4)
730.5 ng/µl
1.84
2.23
50 µl
36.5
91%
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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

 
SpecificationsGenepHlow™ PCR Cleanup KitGenepHlow™ Gel/PCR KitGenepHlow™ DNA Cleanup Maxi KitGenepHlow™ DNA Cleanup Midi KitSmall DNA Fragments Extraction KitLarge DNA Fragments Extraction KitPresto™ 96 Well PCR Cleanup Kit
Cat. #DFC100
DFC300
DFH100
DFH300
DFM010
DFM025
DFI100
DFI300
DF101
DF301
DFL100
DFL300
96DFH04
96DFH10
FormatSpin ColumnSpin ColumnMaxi Spin ColumnMidi Spin ColumnSpin ColumnSilica 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

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