Large DNA Fragments Kit (DFL100, DFL300)

Large DNA Fragments Gel Extraction Kit | Large DNA Fragments PCR Cleanup Kit
Large DNA Fragments Kit (DFL100, DFL300)

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.  

Advantages (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

Specifications
 
Large DNA Fragments Extraction Kit
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

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Catalogue Number
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Large DNA Fragments Extraction Kit
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