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)

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

Related Gel Extraction and PCR Cleanup Products

Product Name	RXNS	Catalogue Number
GenepHlow™ Gel Extraction Kit	100, 300	DFG100/300
GenepHlow™ PCR Cleanup Kit	100, 300	DFC100/300
GenepHlow™ Gel/PCR Kit	100, 300	DFH100/300
GenepHlow™ DNA Cleanup Maxi Kit	10, 25	DFM010/25
Presto™ 96 Well PCR Cleanup Kit	4 x 96, 10 x 96	96DFH04/10
Small DNA Fragments Extraction Kit	100, 300	DF101/301
Large DNA Fragments Extraction Kit	100, 300	DFL100/300
Gel/PCR DNA Fragments Extraction Kit	100, 300	DF100/300
Presto™ 96 Well Gel Extraction Kit	4 x 96, 10 x 96	96DFG04/10
GenepHlow™ DNA Cleanup Midi Kit	100, 300	DFI100/300