This protocol is designed to purify single or doublestranded dna fragments from pcr and other enzymatic reactions see page 8. The weight of the gel is directly proportional to its liquid volume and this is used to determine how much of each buffer to add during the dna isolation step. This kit is specially designed for fast and easy isolation of smaller dna. Add 3 volumes of buffer qg to 1 volume of gel i used the back of 200ul tips. These are available online in convenient and compact pdf format at. Brand thermo scientific qiagen machereynagel promega invitrogen kit and no.
Jenelle meyer 62007 university of wisconsinmadison. Incubate at 3755 c for 510 minutes until the gel slice is completely dissolved2. Qiaex ii handbook qiaex ii agarose gel extraction protocol. Protocol qiaex ii agarose gel extraction protocol this protocol is designed for the extraction of 40bp to 50kb dna fragments from 0. Dna ranging from 70 bp to 10 kb is purified using a simple and fast bindwashelute procedure and an elution volume of 3050. I wanted to know if i can use this kit to purify dna for further transfection in hek293 cells. Qiaquick pcr purification kit protocols 18 using a microcentrifuge 18 using a vacuum manifold 19 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 gel extraction kit for extraction of dna fragments 70 bp 10 kb from. This kit can also be used for dna cleanup from enzymatic reactions see. Dna fragments in a size range of 70 bp to 10 kb can be efficiently recovered. Automated dna cleanup the qiaquick pcr purification kit and qiaquick gel extraction kit can be fully automated on the. Can qiaquick gel extraction kit purified dna be used for cell transfection.
Qiaquick gel extraction kit protocol using a microcentrifuge. Qiagen qiaquick gel extraction kit, 250 rxns, 30 to 50l elution. Dna ladder was run on a 2% agarose gel and 4 different fragment sizes 200 bp, 500 bp, 1 kb, and 5 kb. 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. Qiavac vacuum manifolds 30 handling guidelines for qiavac 24 31 qiavac 6 32. For cleanup of other enzymatic reactions, follow the protocol as described for pcr samples or use the minelute reaction cleanup kit. I cut of my fragment from the agarose and after weighing the weight of my fragments are 460mg and 550mgbut the extraction. Genelute gel extraction kit 70 purifications sigmaaldrich. 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 qiaquick gel extraction kit enables removal of nucleotides, enzymes, salts, agarose, ethidium bromide, and other impurities from samples, ensuring up to 80% recovery of dna see figure high recoveries from gels. This kit can also be used for dna cleanup from enzymatic reactions see page 8. Qiaquick gel extraction kit protocol syracuse university. Qiaex ii gel extraction kit 150 from qiagen selectscience. The gel diffusion buffer, which is not included in the kit, must be prepared prior to beginning the procedure.
The purelink quick gel extraction kit allows you to rapidly and efficiently purify dna fragments from tae or tbe agarose gels of various percentages. Qiaquick pcr purification kit protocols 18 using a microcentrifuge 18 using a vacuum manifold 19 qiaquick nucleotide removal kit protocol 21 qiaquick gel extraction kit protocols 23 using a. Add 3 volumes of buffer qx1 to 1 volume gel 100 mg 100 l. Qiagen qiaex ii gel extraction kit, 150 extractions, 20l elution volume, 5g10l binding. For departmental related matters, please visit the department of molecular, cell, and developmental. Incubate at 3755 c for 510 minutes until the gel slice is completely. I used the qiaquick gel extraction kit and followed the protocol supplied with the kit. The minelute pcr purification kit and minelute gel extraction kit can be fully automated on the qiacube. Gel extraction is a technique used to isolate a desired fragment of intact dna from an agarose gel following agarose gel electrophoresis. Add 3 volumes of buffer qg to 1 volume of gel i used the back of 200ul tips for gel excision and assumed the weight was about 150mg and therefore used 450ul of buffer qg 3. Recoveries of dna fragments in the size range between removed and recovered are not defined. Sigma gel extraction kit is best for dna extraction and purification from gels.
Qiaquick pcr purification kit using a microcentrifuge. Banerjee lab ucla molecular, cell and developmental biology. 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 kit can also be used to purify dna from polyacrylamide gels. Finally, you will want to isolate the dna from the gel. This protocol is designed to extract and purify dna of 70 bp to 10 kb from standard or lowmelt agarose gels in tae or. 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. This is most commonly done with a commercial gel purification kit, such as the qiaquick gel extraction kit. Thermo scientific 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. Qiagen qiaquick gel extraction kit, 50 rxns, 30 to 50l elution. May 24, 2010 gel extraction is a technique used to isolate a desired fragment of intact dna from an agarose gel following agarose gel electrophoresis. The genelute gel extraction kit is designed for isolating dna from agarose gels. 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.
Genelute gel extraction kit na1111 technical bulletin. What is the best gel extraction method for a high dna. The innovative qiacube uses advanced technology to process qiagen spin columns, enabling seamless integration of automated, lowthroughput sample prep into your laboratory workflow. Why is the concentration of dna very low after a gel extraction. For departmental related matters, please visit the department of molecular, cell, and developmental biology website phone. Specific band at 600pb belonging to the b chain is observed. 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 qiaquick gel extraction kit 28704 and 28706. This protocol is designed to extract and purify dna of 70 bp to 10 kb from standard or low melt agarose gels in tae or tbe. Qiaquick gel extraction kit b 0 4 p b 0 4 fragments removed fragments recovered figure 1. Can qiaquick gel extraction kit purified dna be used for. Related products include the qiaquick nucleotide removal kit and qiaquick gel extraction kit. Gel extraction kit and a comparable kit from company q following manufacturers recommended protocols.
Dna can be extracted and purified from agarose gels with. The qiaquick gel extraction kit provides spin columns, buffers, and collection tubes for silicamembranebased purification of dna fragments from gels up to 400 mg slices or enzymatic reactions. Add ethanol 96100% to buffer pe before use see bottle label for volume. 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. The genelute gel extraction kit is designed for the rapid purification of linear and plasmid dna fragments from standard or lowmelting agarose gels. Protocol qiaquick spin handbook 032001 23 qiaquick gel extraction kit protocol using a microcentrifuge this protocol is designed to extract and purify dna of 70 bp qiaquick gel extraction kit for gel the same.
Gel extraction protocol qiaquick gel extraction kit protocol. Qiaquick gel extraction kit 50 from qiagen sample to. For more information, please refer to the qiaquick spin handbook, march 2008. 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.
Up to 400 mg agarose can be processed per spin column. For more information, please refer to the qiaquick spin handbook. Qiaquick gel extraction kit 250 from qiagen selectscience. For purification of dna fragments 40 bp to 50 kb from gels and solutions. Using a microcentrifuge or vacuum manifold, dna ranging from 70 bp to 10 kb is purified from 124 samples. Recoveries of dna fragments in the size range between removed and. For gel extractioncleanup of up to 10 g dna 70 bp to 10 kb from enzymatic reactions. For departmental related matters, please visit the department of molecular, cell, and developmental biology website.
Gel extraction protocol qiaquick gel extraction kit. Dna can be extracted and purified from agarose gels with different melting points in 30 minutes using purelink silica membranebased quick gel extrac. The kit utilizes a proprietary silicabased membrane technology in the form of a convenient spin column. 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.
Electrophoretic analysis of recovered dna from an agarose gel compared to the control. Extraction of dna fragments from polyacrylamide gels qq05. These are available online in convenient and compact pdf format. 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 extractioncleanup of up to 10. For dna fragments 8 kb, following the incubation step, add one additional volume equal to that of the gel slice of. The innovative qiacube uses advanced technology to process qiagen spin columns, enabling. Finally, you will want to isolate the dna from the. Add 3 volumes of adb to each volume of agarose excised from the gel e.
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