Gel/PCR DNA Fragments Extraction Kit

PCR Cleanup Kit
Gel/PCR DNA Fragments Extraction Kit

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 Presto™ Max Gel/PCR 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.

Protocol download

 

 

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
  • Storage: dry at room temperature (15-25ºC) for up to 1 year

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 both the Gel/PCR DNA Fragments Extraction Kit (lane 1) and the equivalent competitor's gel extraction kit (lane 2). The purified DNA from a 50 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder

Test Yield 260/280 260/230 Recovery
Geneaid 6.40 µg/ml 1.83 1.67 92%
6.50 µg/ml 1.84 1.71 94%
Competitor 6.30 µg/ml 1.85 0.45 91%
6.50 µg/ml 1.83 0.51 94%
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 concentration from a 50 µl eluate was confirmed by spectrophotometer and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder

Test DNA Total DNA Recovery
M Control 110 ng/µl 1100 ng ----
1 Test 1 20.8 ng/µl 1040 ng 95%
2 Test 2 20.3 ng/µl 1015 ng 92%
3 Test 3 19.6 ng/µl 980 ng 89%
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
Gel Extraction and PCR Cleanup Kits Specifications
Specifications
 
Cat. #
DFG100
DFG300
DFC100
DFC300
DFH100
DFH300
DF100
DF300
96DFH04
96DFH10
DF101
DF301
DFL100
DFL300
Format
Spin Column
Spin Column
Spin Column
Spin Column
96 Well Plate
Spin Column
Silica Matrix
Binding Capacity 10 µg 10 µg 10 µg 10 µg 10 µg 10 µg 5 µg
Sample Agarose Gel Slice PCR Product Agarose Gel Slice/PCR Product Agarose Gel Slice/PCR Product PCR Product Agarose Gel Slice/PCR Product Agarose Gel Slice/PCR Product
Fragment Size 70 bp-20 kb 70 bp-20 kb 70 bp-20 kb 70 bp-20 kb 70 bp-20 kb 40-200 bp 100 bp-50 kb
pH Indicator Yes Yes Yes N/A N/A N/A Yes
Recovery Up to 90% Up to 95% Up to 95% Up to 95% Up to 95% Up to 95%
Up to 95% (<10 kb)
Up to 80% (10-50 kb)
Elution Volume 20-50 µl 20-50 µl 20-50 µl 20-50 µl 60-80 µl 20-50 µl 10-20 µl
Operation Time 20 minutes 10 minutes <20 minutes <20 minutes 20 minutes <20 minutes <20 minutes
Gel Extraction and PCR Cleanup Products
Product Name
RXNS
Catalogue Number
100, 300
DF100/300
10, 25
DM010/25
100, 300
DF101/301
100, 300
DFL100/300
4 x 96, 10 x 96
96DFH04/10
100, 300
DFG100/300
100, 300
DFC100/300
100, 300
DFH100/300