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Gel/PCR DNA Fragments Kit (DF100, DF300)

The Gel/PCR DNA Fragments Extraction Kit is a PCR Cleanup Kit and Gel Extraction Kit in 1 convenient system. This kit will recover or concentrate 70 bp-20 kb DNA fragments from agarose gel, PCR, or other enzymatic reactions. For processing 10-50 kb DNA fragments, the Large DNA Fragments Extraction Kit is available. For processing 40-200 bp DNA fragments, the Small DNA Fragments Extraction Kit is also available. The Gel/PCR DNA Fragments Extraction Kit uses chaotropic salt to dissolve agarose gel and denature enzymes. DNA fragments in the chaotropic salt are bound by the glass fiber matrix of the gel/pcr 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. 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 such as Fluorescent and Radioactive Sequencing, PCR, Restriction Enzyme Digestion, DNA Labeling and Ligation, Microarray and Transfection.

Quick Protocol

 

 

Advantages (Cat. # DF100, DF300)
  • High Efficiency: up to 90% recovery from agarose gels and up to 95% recovery from PCR products or other enzymatic reactions
  • Convenient: gel extraction kit and PCR cleanup kit in one
  • Only 20 minute gel extraction or 10 minute PCR cleanup
  • Sample Size: up to 300 mg of agarose gel slice or up to 100 μl of PCR products
  • Broad Fragment Size Range: 70 bp–20 kb
  • Spin Columns: glass fiber membrane optimized for both gel extraction and PCR cleanup
  • Elution Volume: 20-50 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

  • DF Buffer
  • W1 Buffer
  • Wash Bufffer
  • Elution Buffer (10 mM Tris-HCl, pH8.5 at 25ºC)
  • DF Columns
  • 2 ml Collection Tubes

Quality Control

The quality of the Gel/PCR 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.

Gel/PCR DNA Fragments Extraction Kit Functional Test Data                

Figure 1. Gel slice DNA fragments ranging from 250 bp-10 kb were extracted using the Gel/PCR DNA Fragments Extraction Kit (total 1.0 µg, 3 replicates). The purified DNA from a 50 µl eluate was analyzed by electrophoresis on a 0.8% agarose gel.

M = Geneaid 1 Kb DNA Ladder

TestDNA Conc.260/280Total DNARecovery
MControl--------1000 ng----
1Test 117.5 ng/µl1.83875 ng87.5%
2Test 216.4 ng/µl1.84820 ng82%
3Test 317.2 ng/µl1.83860 ng86%
Figure 2. PCR product DNA fragments ranging from 250 bp-10 kb were extracted using the Gel/PCR DNA Fragments Extraction Kit (total 1.1 µg, 3 replicates). The purified DNA from a 50 µl eluate was confirmed by spectrophotometer and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder
TestDNA Conc.260/280Total DNARecovery
MControl--------1100 ng----
1Test 120.8 ng/µl1.851040 ng95%
2Test 220.3 ng/µl1.841015 ng92%
3Test 319.6 ng/µl1.84980 ng89%
Figure 3. A 13 kb DNA fragment was purified from agarose gel using the Gel/PCR DNA Fragments Extraction Kit and the equivalent competitors's gel extraction kit. The purified DNA concentration from a 30 µl eluate was confirmed by spectrophotometer and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder
Test
DNA Concentration
Total DNA
Average
Recovery
1
Original 13 kb DNA Fragment (3 µg)
----
----
----
----
2
Geneaid
72.30 ng/µl
2.17 µg
2.31 µg
77%
3
70.10 ng/µl
2.10 µg
4
89.10 ng/µl
1.67 µg
5
Competitor
43.90 ng/µl
1.32 µg
1.27 µg
42%
6
39.80 ng/µl
1.20 µg
7
42.70 ng/µl
1.28 µg
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 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
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 Kit4 x 96, 10 x 9696DFG04/10
GenepHlow™ DNA Cleanup Midi Kit100, 300DFI100/300