Qiaquick pcr purification, qiaquick nucleotide removal, and qiaquick gel extraction. For columns we recommend 4 c storage for periods greater than 1 year. This protocol is designed to extract and purify dna of 70 bp to 10 kb from standard or low. Genelute gel extraction procedure video the genelute gel extraction kit is designed for the rapid purification of 50 bp to 10 kb linear dna fragments and plasmids from standard or lowmelting agarose gels. I put in 1ug and i am getting back about 10nul but, my 260280s are jumping all over the place, as is my concentrations. Qiaquick gel extraction kit b 0 4 p b 0 4 fragments removed fragments recovered figure 1. For uptodate licensing information and productspecific disclaimers, see the respective qiagen kit handbook or user. This protocol is designed to extract and purify dna of 70 bp. If the purified dna is to be analyzed on a gel, add 1 volume of loading dye to. It eliminates resin manipulation, toxic phenol chloroform extractions, and ethanol precipitations.
For gel extractioncleanup of up to 10 g dna 70 bp to 10 kb from enzymatic reactions. The innovative qiacube uses advanced technology to process qiagen spin columns, enabling seamless integration of automated, lowthroughput sample prep into your laboratory workflow. Solutions that contain ethanol, isopropanol or mops should be sterilized by filtration only. Qiaquick gel extraction kit protocol using a microcentrifuge. Purelink quick gel extraction and pcr purification combo kit. Monarch dna gel extraction kit reproducibly recovers dna over a broad range of molecular weights. Qiaamp dna mini kit and qiaamp dna blood mini kit handbook. They are not to be used for human diagnostic or drug purposes or to be administered to humans unless expressly cleared for that purpose by the food and drug administration in the usa. Qiaquick gel extraction and pcr purification kit from qiagen. Dna adsorbs to the dneasy membrane in the presence of high concentrations. The binding buffers in minelute spin kits provide the correct salt concentration. The most versatile benchtop automation in the lab for consistent extraction and purification of dna, rna, protein, and cells.
Qiaex ii agarose gel extraction protocol this protocol is designed for the extraction of 40bp to 50kb dna fragments from 0. Qiavac vacuum manifolds 30 handling guidelines for qiavac 6 31 qiavac 24 32 ordering information 33 qiagen companies and distributors 35 qiaquick spin handbook 072002 3. Lyophilized qiagen protease can be stored at room temperature 1525c for up to. Visual inspection of rna integrity by agarose gel electrophoresis. Automated extraction of gdna from buffy coat samples. For more information, please refer to the qiaquick spin handbook, march 2008. Recoveries of dna fragments in the size range between removed and recovered are not defined. Kit for total dna isolation from agarose gels agilent. Qiagen plasmid purification handbook harvard university. Dna ladder was run on a 2% agarose gel and 4 different fragment sizes 200 bp, 500 bp, 1 kb, and 5 kb were recovered using omega bioteks e.
The genejet gel extraction kit is designed for rapid and efficient purification of dna fragments from standard or lowmelting point agarose gels run in either tae or tbe buffer. For purification of dna fragments 40 bp to 50 kb from gels and solutions. Rapid regeneration and reuse of silica columns from pcr. Banerjee lab ucla molecular, cell and developmental biology. The zymoclean gel dna recovery kits are gel dna extraction kits that provide rapid purification of highquality dna from taetbebuffered agarose gels. Gel extraction kit and a comparable kit from company q following manufacturers recommended protocols. Add ethanol 96100% to buffer pe before use see bottle label for volume. The minelute pcr purification kit and minelute gel extraction kit can be fully automated on the qiacube. Qiaquick gel extraction kit 40 b 40 bp fragments removed fragments recovered recoveries of dna fragments in the size range between removed and recovered are not defined. A mixture of 7 dna fragments ranging from 10 kb down to 0. The thermo scientific genejet gel extraction kit should be stored at room temperature 1525 c. Qiagen qiaquick gel extraction kit 28704 and 28706.
Isolate a suitable piece of tissue and place in a uvcrosslinked 1. Load more dna on the gel and take care that you excise the gel. If you get double bands consistently, do a gel extraction see below. Forceps are to be sterilized in fine science tools heat block at. Qiagen genomictips are gravityflow, anionexchange tips that enable purification of dna of up to 150 kb from a wide range of sample types. These gel dna extraction kits feature zymospin technology to yield highquality, purified dna in just minutes. Qiagen plasmid purification handbook 082003 5 kit contents. We use qiagen gel extraction kit and we have not faced such problem with concentration of dna. Qiaex ii handbook qiaex ii agarose gel extraction protocol.
For maximum convenience and value, columns and buffers are also available separately. Qiaex ii gel extraction kit 150 from qiagen selectscience. Rneasy minihandbook rneasy mini kit for purification of total rna from animal cells, animal tissues, bacteria, and yeast, and for rna cleanup rneasy protect mini kit for immediate stabilization of rna in harvested animal tissues and subsequent total rna purification rneasy plant mini kit for purification of total rna from plants and. All pcr products were excised from the gel and purified using qiaquick gel extraction kit qiagen, germany. Dna fragments of interest are extracted by solubilizing slices of an agarose gel. Minelute gel extraction kit 50 from qiagen selectscience.
Electrophoretic analysis of recovered dna from an agarose gel compared to the control. Qiagen qiaquick gel extraction kit, 50 rxns, 30 to 50l elution volume, 10g binding capacity, dna sample, tube format, silica technology, manual processing, 70 bp to 10 kb fragment, fast and convenient procedure, for gel extractioncleanup of up to 10g dna 70. Qiagen plasmid purification handbook 082003 9 product use limitations qiagen, qiafilter, and endofree plasmid kits are developed, designed and sold for. The kingfisher system technology provides an efficient workflow solution. Qiagen qiaex ii gel extraction kit, 150 extractions, 20l elution volume, 5g10l binding capacity, dna sample, tube format, silica technology, manual processing, 40 bp to 50 kb fragment, ideal for restriction digestion, labeling, ligation, pcr, for purification of dna fragments. Qiaquick nucleotide removal kit protocol 21 qiaquick gel extraction kit protocols 23 using a microcentrifuge 23 using a vacuum manifold 25 troubleshooting guide 28 appendix. Qiaquick pcr purification kit protocol using a microcentrifuge this protocol is designed to purify single or doublestranded dna fragments from pcr and other enzymatic reactions see page 8. Dnafragments yes yes yes yes oligonucleotides no yes no no ligation yes yes yes yes nicktranslation yes yes yes yes pcr yes. Formulas for qiagen kit buffers genesee scientific. For departmental related matters, please visit the department of molecular, cell, and developmental biology website. For cleanup of other enzymatic reactions, follow the protocol as described for pcr samples or use the minelute reaction cleanup kit.
Each fragment was manually excised from the agarose gel and processed using the monarch dna gel extraction kit. Qiaquick gel extraction kit protocols 23 using a microcentrifuge 23 using a vacuum manifold 25 troubleshooting guide 28 appendix. Qiavac vacuum manifolds 30 handling guidelines for qiavac 24 31 qiavac 6 32 ordering information 33 qiagen companies and distributors 35 qiaquick spin handbook 032001 3. Download download qiagen 28704 manual read online read online qiagen 28704 manual qiaquick pcr purification kit protocol gel extraction principle qiagen buffer qg qiagen gel extraction low yield qiaex ii gel extraction kit qiagen gel extraction troubleshooting gel extraction protocol without kit qiagen kit 28704 protocol this protocol is designed to extract and purify. Poor yield with gel extraction kit for about the umpteenth time now, i have gotten poor yield isolating my cut fragment from a 1% agarose gel. Automated extraction of gdna from buffy coat samples preserved. Disposable columns from qiagen pcr purification and gel extraction kits can be repeatedly regenerated and reused a minimum of five times. Qiaquick gel extraction kit protocol using a microcentrifuge chen. Minelute handbook 062004 11 adsorption to minelute membrane salt and ph dependence the minelute silicagel membrane is uniquely adapted to isolate dna from both aqueous solutions and agarose gels, and up to 5 g dna can bind to each minelute column. The genelute gel extraction kit combines silicabinding technology with the convenience of a spin or vacuum column format.
I have one from qiagen but i only recover about 50 % of my dna. This kit can also be used for dna cleanup from enzymatic reactions see page 8. Qiagen s qiaquick gel extraction kit is highly effective for extracting high purity, doublestranded dna from either pcr reactions or agarose gels following gel electrophoresis. For gel extraction or cleanup of dna 70 bp 10 kb from enzymatic reactions. Any precipitate that forms in the buffers during storage can be redissolved by incubating briefly at 37 c, then cooling to room temperature before use.
Using the small gel rig, pour a 1% low melting point agarose gel 0. Qiagen qiaquick gel extraction kit, 250 rxns, 30 to 50l elution volume, 10g binding capacity, dna sample, tube format, silica technology, manual processing, 70 bp to 10 kb fragment, fast and convenient procedure, for gel extraction cleanup of up to. The qiaquick pcr purification kit and qiaquick gel extraction kit can be fully automated on the qiacube. Qiagen qiaquick gel extraction kit, 50 rxns, 30 to 50l elution volume, 10g binding capacity, dna sample, tube format, silica technology, manual processing, 70 bp to 10 kb fragment, fast and convenient procedure, for gel extraction.
L of qg buffer were added to 1 volume 100 mg gel slice of gel containing excised dna fragment. Marker qiagen kit brain liver heart purelink kit qiagen kit purelink kit qiagen kit purelink kit marker figure 5. Quality control the quality of the presto 96 well gel extraction kit is tested on a lottolot basis by purifying dna fragments of various sizes from agarose gel. The gel solubilization solution dissolves an agarose gel slice from gels run in. Thermo scientific genejet gel extraction kit thermo fisher.
Why is the concentration of dna very low after a gel extraction. Agarose gel analysis of the purification procedure 60. For departmental related matters, please visit the department of molecular, cell, and developmental biology website phone. As assessed by agarose gel, the dna extracted from each sample had a molecular weight equal to or greater than 23 kb figure 2. Automated dna cleanup the qiaquick pcr purification kit and qiaquick gel extraction kit can be fully automated on the. The axyprep dna gel extraction kit employs optimized reagents in combination with a convenient miniprep column to purify dna fragments from either tae or tbe agarose gels regular and lowmelt. This protocol is designed for purification of total dna from grampositive bacteria. Kit is a universal kit for dna extraction and purification from a wide variety of cell types, including blood, cells, tissues and toughtolyse samples including bacteria and yeast. Qiagen qiaquick gel extraction kit, 50 rxns, 30 to 50l elution volume, 10g binding capacity, dna sample, tube format, silica technology, manual processing, 70 bp to 10 kb fragment, fast and convenient procedure, for gel extraction cleanup of up to 10g dna 70. Qiaquick gel extraction kit protocol using a microcentrifuge this protocol is designed to extract and purify dna of 70 bp to 10 kb from standard or lowmelt agarose gels in tae or tbe buffer. The kit was tested in the extraction of dna fragments from an agarose gel according to the protocol described in the manual. If you get very faint bands, clean as usual using ampure, then concentrate product see below. Qiaquick gel extraction kit 250 from qiagen selectscience.
For gel extraction cleanup of up to 10 g dna 70 bp to 10 kb from enzymatic reactions. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. Automated purification and extraction thermo fisher. Qiaquick gel extraction kit qiagen, 28704 or qiaquick pcr purification kit qiagen, 28104, see below. The purelink quick gel extraction and pcr purification combo kit is designed to purify dna fragments from agarose gels. They can be stored for at least 2 years without showing any reduction in performance, capacity, or quality of separation. The qiaquick gel extraction kit provides spin columns, buffers, and collection tubes for silicamembranebased. The kit utilizes a proprietary silicabased membrane technology in the form of.
Using a microcentrifuge or vacuum manifold, dna ranging from 70 bp to 10 kb is purified from 124 samples. Explore how strataprep dna gel extraction kits allow rapid, highyield isolation of pure dna from agarose gels. Download download qiagen 28704 manual read online read online qiagen 28704 manual qiaquick pcr purification kit protocol gel extraction principle qiagen buffer qg qiagen gel extraction low yield qiaex ii gel extraction kit qiagen gel extraction troubleshooting gel extraction protocol without kit qiagen kit 28704 protocol this protocol is designed to extract and purify dna of 70 bp to 10. Product information thermo scientific genejet gel extraction kit. The protocol describes the preliminary harvesting of bacteria and incubation with lysozyme to lyse their cell walls before dna purification. Qiaquick gel extraction kit the qiaquick gel extraction kit cat.
Up to 400 mg agarose can be processed per spin column. Monarch nucleic acid purification kits are optimized for maximum performance and minimal environmental impact. Qiaquick gel extraction kit 50 from qiagen sample to. All samples were from 30 mg tissue input and showed a high level of rna integrity. Dna fragments in a size range of 70 bp to 10 kb can be efficiently recovered. Dna ranging from 70 bp to 10 kb is purified using a simple and fast bindwashelute procedure and an elution volume of 3050.
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