Can i use the gel loading dye, purple with the qiaquick gel extraction kit. Qiaquick gel extraction kit 10 16 2 qiaquick spin032008. Dna ranging from 70 bp to 10 kb is purified using a simple and fast bindwashelute procedure and an elution volume of 3050. Qiaquick kits contain a silica membrane assembly in a spin column for binding of dna in highsalt buffer and elution with lowsalt buffer or water. Qiaquick pcr purification kit protocol fragments ranging from 100 bp to 10 kb are purified from primers, nucleotides, polymerases, and salts using qiaquick spin columns in a microcentrifuge. I cut of my fragment from the agarose and after weighing the weight of my fragments are 460mg and 550mgbut the extraction kit says for the slice of more than 400mg we should use more than one qia column. For gel extraction or cleanup of dna 70 bp 10 kb from enzymatic reactions.
Qiagen qiaquick gel extraction kit 28704 and 28706. 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. Pcr purification qiaquick kit protocol this protocol is designed to purify single or doublestranded dna fragments from pcr. The extraction method avoids the use of liquid nitrogen, requires minimal amount 50 mg of plant tissue, and allows the recovery of 0. This kit can also be used for dna cleanup from enzymatic.
Qiaquick gel extraction kit the qiaquick gel extraction kit cat. For purification of dna fragments 40 bp to 50 kb from gels and solutions. Qiaquick gel extraction kit from qiagen biocompare. This protocol is designed to purify single or doublestranded dna fragments from pcr and other enzymatic reactions see page 8. Freezethaw method of extraction is a commonly used advantageous dna recovering method which will supports the common laboratory facilities. Can i use the gel loading dye, purple with the qiaquick. Qiaquick gel extraction kit protocol using a microcentrifuge. How to quality control check your rna samples bitesize bio. Recovery efficiency of dna fragments ranging from 70 bp to 10 kb in size is up to 80%. Fragments ranging from 100 bp to 10 kb are purified from primers, nucleotides, polymerases, and salts using qiaquick spin columns in a microcentrifuge. Qiagen qiaquick pcr purification kit, 250 rxns, 10g binding capacity, 30l elution volume, tube format, manual processing, silica technology, 100 bp to 10 kb fragment, 40mers fragments removed, ideal for sequencing, microarray analysis, ligation and transformation, restriction. The overall quality of an rna preparation may be assessed by electrophoresis on a denaturing agarose gel. Qiaquick spin handbook en print bookmark share pdf 814kb.
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. Qiaquick spin handbook qiaquick pcr purification kit for purification of pcr products, 100 bp to 10 kb qiaquick nucleotide removal kit for oligonucleotide 1740mers and dna 40 bp to 10 kb cleanup from enzymatic reactions qiaquick gel extraction kit for gel extraction or cleanup of dna 70 bp to 10 kb from enzymatic reactions second. As reedms said, make sure its at the right ph, but this is only likely to be a problem for old stock. Qiaquick spin handbook 032008 19 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 qiaquick gel extraction kit provides spin columns, buffers, and collection tubes for silicamembranebased. I put in 1ug and i am getting back about 10nul but, my 260280s are jumping all over the place, as is my concentrations. For purification of up to 10 micro g pcr products, 100 bp to 10 kb. It is easy to underestimate the problems that residual agarose or salt can cause in some downstream applications. Qiaquick gel extraction kit 250 from qiagen selectscience. I used the qiaquick gel extraction kit and followed the protocol supplied with the kit. Qiaquick 96 pcr purification kit, or qiaquick 96 pcr biorobot kit should be used. Qiaquick pcr purification, qiaquick nucleotide removal, and qiaquick gel extraction.
A denaturing gel system is suggested because most rna forms extensive secondary structure via intramolecular base pairing, and this prevents it from migrating strictly according to its size. It is very simple and easy to perform with good yield. 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 10micro g. 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. A260230 readings post gel extraction general lab techniques. Run the gel for 2,5 hours in 0,5x tbe at 1 macm at the start the voltage is about 120 v, during the course of electrophoresis the voltage increases to 200 v. All centrifuge steps are at max speed 15000 rpm in a tabletopmicrocentrifuge. 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. Dna can be extracted and purified from agarose gels with different melting points in 30 minutes using purelink silica. If purified dna is to be analyzed on a gel, add 1 volume of loading dye to 5 volumes of purified dna. 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 to 10 kb from standard or lowmelt agarose gels in tae or tbe buffer. Qiaprep miniprep handbook 122006 3 contents kit contents 4 storage 7 quality control 7 product use limitations 7. You can anyone suggest me how to proceed with my extraction.
Rna extraction products thermo fisher scientific in. Qiaquick gel extraction kit protocol using a microcentrifuge chen. Regardless of 260230 or 260280, it is always optimal to get at least 1. 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. The purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide, and other impurities from dna samples. Abstract we present a protocol for construction of tunable crispr interference tcrispri strains for escherichia coli. Using a microcentrifuge or vacuum manifold, dna ranging from 70 bp to 10 kb is purified from 124 samples. This should only be of concern if the dna band you are extracting migrates where the dye migrates. Do not add bleach or acidic solutions directly to the samplepreparation.
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. Gel extraction qiagen qiaquick gel extraction kit materials. The purelink quick gel extraction kit allows you to rapidly and efficiently purify dna fragments from tae or tbe agarose gels of various percentages. Rneasy minelute cleanup handbook 102010 7 introduction the rneasy minelute cleanup kit uses novel technology to purify and concentrate rna. Most of the molecular biology laboratories use low melting point agarose for the separation of dna from agarose. Qiaex ii gel extraction kit en print bookmark share pdf 48kb english format file size language download get adobe reader contact qiagen. The qiaquick pcr purification kit provides spin columns, buffers, and collection tubes for silicamembranebased purification of pcr products 100 bp.
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. Apr 08, 2008 if you really want to go for the maximum concentration after gel extraction, it could be worth considering zymos dna gel extraction kit, as this allows elution with only 6 microlitres. Qiaquick spin handbook 072002 23 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. Dna of up to 10 kb is purified using a simple and fast bindwashelute procedure and an elution volume of 3050. Qiaquick gel extraction kit protocol syracuse university. This protocol is designed to extract and purify dna. Qiaquick gel extraction kit 50, 28704 from qiagen labsave.
Some of these familiar products include ambion cellstoct kits, trizol reagents, purelink purification kits, magmax extraction kits, mirvana isolation kits, and dynabeads mrna isolation technologies. I always carry out the gel extraction optional steps and they have always gone fine. After page, transfer the gel to a double layer of whatman paper and dry the gel on a gel dryer for 45. Please keep in mind that gel loading dye, purple contains a redcolored dye which will create an orange color similar to an indication that the ph has changed. The qiaex ii gel extraction kit provides qiaex ii suspension together with binding and wash buffers, and a comprehensive handbook. Phenol extraction and ethanol precipitation are not required, and highquality plasmid dna is eluted in. For gel extractioncleanup of up to 10 micro g dna 70 bp to 10 kb from enzymatic reactions. Qiaquick pcr purification kit 250, 28106 from qiagen. Gelpcr dna fragments kit df100, df300 genephlow gelpcr kit dfh100, dfh300 large dna fragments kit dfl100, dfl300. For uptodate licensing information and productspecific disclaimers, see the respective qiagen kit handbook or user manual. Mar 30, 2007 we use qiagens qiaquick gel extraction kit for this purpose. We generally purify pcr products from 1% agarose gels, but you can purify from up to 2% agarose, even more than 2% agarose by making small modifications to the.
Up to 400 mg agarose can be processed per spin column. For departmental related matters, please visit the department of. This kit may also may used for restriction enzyme digest reactions,ligation reactions. See qiaex ii gel extraction kit protocol for desalting and concentrating dna from solutions. Use gel extraction tips to extract the dna band from the gel. Mix the solution by pipetting up and down before loading the gel. Isolate a suitable piece of tissue and place in a uvcrosslinked 1. It totally eliminate slurry carryover problem as encountered using silica beads or glass milk based methods. 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. Jul 22, 2009 i use the qiagen gel extraction kit all the time and have never encountered such a low 260230. Gel containing dna to extract gel extraction tips buffer qg 3 volumes gel extract 100 mg 100. Qiaex ii gel extraction kit 150 from qiagen selectscience. For departmental related matters, please visit the department of molecular, cell, and developmental biology website phone.
Dna fragments in a size range of 70 bp to 10 kb can be efficiently recovered. Rneasy minihandbook rneasy mini kit for purification of total rna from animal cells, animal tissues, bacteria, and yeast, and for rna cleanup. The kit is suitable to extract and purify dna of 70 bp to 10 kb from standard or low melt agarose gels in tae or tbe. The qiaquick pcr purification kit, qiaquick nucleotide removal kit, qiaquick gel. How to improve the yield of dna after gel purification. Most of the molecular biology laboratories use low melting point. Four microliters of either egel extract or qiaquick gel puri. The tcrispri system alleviates most of the known problems of plasmidbased expression methods, and can be immediately used to construct libraries of sgrnas that can complement the keio collection by targeting both essential and nonessential genes. Forceps are to be sterilized in fine science tools heat block at. Our rna extraction products include organic, silica spin columns, crude lysate, and magnetic beads. Banerjee lab ucla molecular, cell and developmental biology.
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