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Large DNA Fragments Kit (DFL100, DFL300)

Large DNA Fragments Kit (DFL100, DFL300)

Large DNA Fragments Kit (DFL100, DFL300)

DNA Extraction/Gel Extraction

 

The Large DNA Fragments Extraction Kit is a PCR Cleanup Kit and Gel Extraction Kit in 1 convenient system optimized to recover or concentrate a broad range of DNA fragments (100 bp-50 kb) from agarose gel, PCR, or other enzymatic reactions. The Large DNA Fragments Extraction Kit uses a concentrated sodium buffer to dissolve agarose gel and denature enzymes. DNA fragments are then bound by silica beads. 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. Salts, enzymes and unincorporated nucleotides can be effectively removed from the reaction mixture without phenol extraction or alcohol precipitation. Typically, recoveries are up to 80% for Gel Extraction and up to 95% for PCR Cleanup. The entire procedure can be completed in 20 minutes and DNA can be eluted in (10-20 μl) of Elution Buffer to obtain a high concentration of DNA products. The purified DNA is ready for use in PCR, Fluorescent or Radioactive Sequencing, Restriction Enzyme Digestion, DNA Labeling and Ligation.

 

DESCRIPTION

Specifications (Cat. # DFL100, DFL300)

  • Purify large DNA fragments within 20 minutes!
  • High Recovery of large DNA fragments: up to 80% (10-50 kb), up to 95% (<10 kb)
  • Optimized for 100 bp-50 kb DNA fragment recovery
  • Sample Size: up to 250 mg (agarose gel slice) or up to 100 μl (PCR products)
  • Elution volume: 10-20 μl for highly concentrated DNA
  • Modified silica bead protocol
  • Process both TAE and TBE agarose gel
  • Convenient: DF2 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
  • Storage: dry at room temperature (15-25ºC)

APPLICATIONS

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

COMPONENTS

  • Presto™ Max Suspension
  • DF2 Buffer
  • 3M Sodium Acetate (pH5.0)
  • Wash Buffer
  • Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)

QUALITY CONTROL

The quality of the Large DNA Fragments Extraction 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 and analyzed by electrophoresis.

Large DNA Fragments Extraction Kit Functional Test Data

Figure 1 .Genomic DNA was extracted from 1x106 Hela cells using a traditional phenol-chloroform method combined with ethanol precipitation. 2 µg of extracted genomic DNA (30 kb) was then purified using the Large DNA Fragments Extraction Kit. The quality and concentration of genomic DNA was significantly increased following the purification process.

M = Geneaid 1 Kb DNA Ladder

 

Sample

ng/µl

260/280

260/230

Yield (µg)

Recovery

1 - Unpurified 30 kb gDNA

20

1.67

0.92

2.0

-----------

2 - Purified 30 kb gDNA

64.8

1.89

1.74

1.3

65%

3 - Purified 30 kb gDNA

61.5

1.87

1.75

1.23

61.5%

Figure 2 .A 13 kb DNA fragment (lane 1, original 13 kb DNA fragment, 1,000 ng) was purified from agarose gel using the Large DNA Fragments Extraction Kit. 20 µl eluates were analyzed by electrophoresis on a 1% agarose gel.

M = Geneaid 1 Kb DNA Ladder

 

Sample

ng/µl

Final Volume

Yield (ng)

Recovery

Test 1

39.70

20.00 µl

794.00

79.40 %

Test 2

40.80

20.00 µl

816.00

81.60%

Large DNA Fragments Extraction Kit Sequencing Data

Figure 3 .Sequencing data of DNA purified using the Presto™ Max Gel/PCR Kit

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

 

DOCUMENTS

 

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