DNA Cleanup Midi Kit (DFI100, DFI300)

GenepHlow™ DNA Cleanup Midi Kits were designed to recover or concentrate DNA fragments from PCR or other enzymatic reactions. This DNA cleanup kit includes a pH indicator as an optional addition to the PB binding buffer to ensure optimal pH and facilitate DNA binding. If pH exceeds the optimal level (>7.5), the color of the solution will appear purple instead of yellow. 3M Sodium Acetate (pH5.0) which is included with the kit, can then be added to the solution to adjust pH and return the color to yellow. Chaotropic salt is used to denature enzymes while DNA fragments are bound by the glass fiber matrix of the 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. The pH indicator, 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. 

Advantages (Cat. # DFI100, DFI300)

 

GenepHlow™ DNA Cleanup Midi Kit Functional Test Data 

Figure 1. 40 µg of pbluescript plasmid DNA was digested by Hind III endonuclease at 37ºC for 1 hour. Following digestion, the linear DNA fragment was purified using the GenepHlow™ DNA Cleanup Midi Kit. The unpurified DNA fragment (lane 1) and 3 replications of 5 µl purified linear DNA fragments (lanes 2-4) were loaded on 0.8% agarose gel. M = Geneaid 1 Kb DNA ladder Sample ng/µl 260/280 260/230 Elution Yield (µg) Recovery 2 - Purified DNA 731.4 ng/µl 1.84 2.28 50 µl 36.6 92% 3 - Purified DNA 724.8 ng/µl 1.83 2.24 50 µl 36.2 91% 4 - Purified DNA 730.5 ng/µl 1.84 2.23 50 µl 36.5 91%

Optimal pH   pH Too High

Figure 2. A pH indicator is included as an optional addition to the PB binding buffer to ensure optimal pH and facilitate DNA binding. If pH exceeds the optimal level (>7.5), the color of the solution will appear purple instead of yellow. 3M Sodium Acetate (pH5.0), which is included with the kit, can then be added to the solution to adjust pH and return the color to yellow.

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