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제품 설명
Large DNA Fragments Kit (DFL100, DFL300)
제품 번호
DFL100
제품 특징
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
| Specifications | GenepHlow™ PCR Cleanup Kit | GenepHlow™ Gel/PCR Kit | GenepHlow™ DNA Cleanup Maxi Kit | GenepHlow™ DNA Cleanup Midi Kit | Small DNA Fragments Extraction Kit | 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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