Pcr purification qiaquick kit protocol this protocol is designed to purify single or doublestranded dna fragments from pcr. The minelute reaction cleanup kit provides spin columns, buffers, and collection tubes for silica membranebased. For gel extraction of up to 5 micro g dna fragments 70 bp to 4 kb in low elution volumes. The handbook supplied with your qiasymphony important kit will tell you which protocol you. The protocol describes the preliminary harvesting of bacteria and incubation with lysozyme to lyse their cell walls before dna purification. For emcv skip overnight culture, go directly from plate into 100 mls of lb. Dna adsorbs to the dneasy membrane in the presence of high concentrations of chaotropic salt, which remove. In accordance with qiagens isocertified quality management system. Minelute columns are designed to give high endconcentrations of purified dna fragments for subsequent reactions. Jenkins lab mhc class ii tetramer production protocol 010816.
If you need to isolate your band of interest from other bands in the digest, proceed to step 6. For cleanup of other enzymatic reactions, follow the protocol as described for pcr samples or use the minelute reaction cleanup kit. View and download qiagen qiasymphony sp operating manual online. Qiagen distributors please see the last page for contact information for your local qiagen distributor. For purification of lowcopy plasmids and cosmids, large plasmids 10. 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. Qiagen qiaquick gel extraction kit, 50 rxns, 30 to 50l elution volume, 10g binding capacity, dna sample, tube format, silica technology, manual. Applicationsfor gel extractioncleanup of up to 10 g. Washthecolumnwith4resinvolumeofdenaturingbindingbufferbyresuspendingtheresin androckingfortwominutes. Principle and procedure qiagen plasmid purification protocols are based on a modified alkaline lysis procedure, followed by binding of plasmid dna to qiagen anionexchange resin under. Qiaquick pcr purification kit 250, 28106 from qiagen labsave. Under these conditions, qiaquick spin kits can be stored for up to 12 months without showing any reduction in performance and quality. Pcr purification kit qiagen protocol qiaquick pcr purification kit. Gel extraction qiagen qiaquick gel extraction kit materials.
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. Using a microcentrifuge or vacuum manifold, dna ranging from 70 bp to 10 kb is purified from 124 samples. Qiavac vacuum manifolds 30 handling guidelines for. This protocol describes the preparation of libraries of chromatinimmunoprecipitated dna in a format compatible with the illuminas cluster amplification and sequencing platforms. Qiagen minelute gel extraction kit, 50 minelute spin columns, 5g binding capacity, 10l elution volume, tube format, gel extraction technology, 70 bp to 4 kb fragment size, manual processing, dna sample, fast procedure and easy handling, high, reproducible. This product is not intended for the diagnosis, prevention, or treatment of a disease. For uptodate licensing information and productspecific disclaimers, see the respective qiagen kit handbook or user manual.
Australia qiagen pty ltd po box 25 clifton hill victoria 3068 abn 75 072 382 944 orders 0394893666 fax 0394893888 technical 1800243066 canada qiagen inc. The following protocol for sds removal was developed for cytoplasmic intermediate filaments, and we successfully applied it to lamin a. Banerjee lab ucla molecular, cell and developmental biology. Qiagen plasmid purification handbook harvard university. Allowseveralweeks2k3weeksusuallyforeachsinglecelltogrowintoavisiblecolony, inspectcellseveryweekandchange. Qiacube follows the same steps as the manual procedure i. 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. Background information, protocols, hints, and tips are provided for purification and analysis of plasmid dna, genomic dna, rna, and proteins, as well as recipes for buffers and solutions. Initiate starter culture in the morning from an isolated colony from an agar plate with the appropriate selective antibiotic.
Qiagen kit handbooks and user manuals are available at. 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. The kit is suitable to extract and purify dna of 70 bp to 10 kb from. Sample preparation for chip kit protocol version 1. If the instrument becomes electrically unsafe, prevent other personnel from operating it, and contact qiagen technical services. Qiaquick gel extraction kit protocol using a microcentrifuge chen. 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. For departmental related matters, please visit the department of molecular, cell, and developmental biology website. We use qiagens qiaquick gel extraction kit for this purpose.
The ultrafast purification protocol, based on the remarkable selectivity of patented qiagen resin, allows the. Qiaquick kits contain a silica membrane assembly for binding of dna in highsalt buffer and elution with lowsalt buffer or water. Qiaquick gel extraction kit protocol using a microcentrifuge. Qiaquick spin handbook for qiaquick pcr purification kit qiaquick nucleotide removal kit. In the united states, the centers for disease control and prevention cdc protocol requires the use of the qiagen qiaamp dsp viral rna mini kit. For more information, please refer to the qiaquick spin handbook, march 2008. Our advanced, highquality products and services ensure success from sample to result. The rnasefree dnase set is intended for molecular biology applications. Qiagen guarantees the performance of all products in the manner described in our product literature. Safety information when working with chemicals, always wear a suitable lab coat, disposable. Gel containing dna to extract gel extraction tips buffer qg 3 volumes gel extract 100 mg 100. Qiagen is the leading provider of innovative sample and assay technologies. Qiaquick pcr purification kit 28104 pdf click here qiagen 28104 per the qiaquick protocol and elute with 32 il eb warmed to 55 c.
This protocol is designed to purify single or doublestranded dna fragments from pcr and other enzymatic reactions see page 8. Qiaquick protocols 18 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. 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 steps as the manual qg and leads to inefficient dna binding. Hispeed plasmid purification handbook harvard university. Forceps are to be sterilized in fine science tools heat block at. Qiaquick gel extraction kit from qiagen biocompare product. Australia qiagen pty ltd po box 25 clifton hill victoria 3068 abn 75 072 382 944 orders 0394893666 fax 0394893888 technical 0394893666 canada qiagen inc. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol.
Qiaquick spin handbook qiaquickpcrpurificationkit forpurificationofpcrproducts,100bpto10kb qiaquicknucleotideremovalkit foroligonucleotide17 40mersanddna 40bpto10kbcleanupfromenzymaticreactions. Qiagen allprep dnarna ffpe kit protocol pugh lab printed. In this handbook, different protocols are provided for different starting materials. Many companies sell kits for purifying dna from enzymatic reactions for example, qiagen sells the qiaquick pcr purification kit or qiaquick gel extraction kit. Purification of pcr products for sequencing the following protocols for preparing good quality dna templates for sequencing are recommended by applied biosystems and can be found online in their. Purification of plasmid dna for microinjection 1 prepare plasmid from bacteria using your preferred method alkaline lysis, lysozymetriton or other methods, but be sure to perform 2 cscl gradients. For gel extractioncleanup of up to 10 micro g dna 70 bp to 10 kb from enzymatic reactions. Isolate a suitable piece of tissue and place in a uvcrosslinked 1. Minelute gel extraction kit 50, 28604 from qiagen labsave. The primary assay employed is a reverse transcription quantitative polymerase chain reaction rtqpcr assay that requires the use of an rna extraction kit 22,33. Qiagen qiaquick gel extraction kit 28704 and 28706. 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. A protocol is a set of instructions that allows the qiasymphony sp to perform a molecular biology application.
Qiaquick gel extraction kit the qiaquick gel extraction kit cat. Also note that phenol kills injected embryos so an ethanol precipitation and ethanol wash must follow all phenol extractions. Automated low to moderatethroughput for dna purification 20 f. These are available online in convenient and compact pdf format. It has been stored under unfavorable conditions for a prolonged period. Once the sample is bound to the membrane, the protocols are similar see flowchart, next page. Mar 30, 2007 extraction of pure dna from agarose is essential for many downstream assays including cloning and radioisotopic or fluorescent dna sequencing. Should any product fail to perform satisfactorily due to any reason other than misuse, qiagen will replace it free of charge or refund the purchase price. 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 to 10 kb from standard or lowmelt agarose gels in tae or tbe buffer.
Otherwise, purify your dna and proceed to step 10 ligation. Our technical service departments are staffed by experienced scientists with extensive practical and theoretical expertise in molecular biology and the use of qiagen prod ucts. For purification of up to 10 micro g pcr products, 100 bp to 10 kb. The qiagen bench guide is designed to help you with your laboratory work. The purchaser must determine the suitability of the product for its particular use. This protocol is designed for purification of total dna from grampositive bacteria. Different net proteins and fragments will bind sds. Qiaquick gel extraction kit 50 from qiagen sample to insight. Qiaprep spin midimaxiprep kit protocol before staring make sure that rnase a has been added to p1 buffer. Jenkins lab 1 1272016 jenkins lab mhc class ii tetramer production protocol 0108 16 check for updates at. Quickstart protocol july 2015 qiaquick gel extraction kit the qiaquick gel extraction kit cat. The purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide, and other impurities from dna samples see figure high recoveries from gels. Qiaquick spin kits should be stored dry at room temperature 1525c.
Dna adsorbs to the dneasy membrane in the presence of high concentrations. For departmental related matters, please visit the department of. Qiaquick gel extraction kit 50, 28704 from qiagen labsave. Up to 400 mg agarose can be processed per spin column. Qiagen qiasymphony sp operating manual pdf download. Qiagen plasmid purification handbook 082003 9 product use limitations qiagen, qiafilter, and endofree plasmid kits are developed, designed and sold for research purposes only. The procedure can be fully automated on the qiacube connect. 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. Purification of total dna from animal tissues spincolumn protocol this protocol is designed for purification of total dna from animal tissues, including rodent tails. Qiagen is the leading provider of innovative sample and assay technologies, enabling. Under these conditions, if no expiration date is mentioned on the kit label, qiaquick kits can be stored for at least. Fragments ranging from 100 bp to 10 kb are purified from primers, nucleotides, polymerases, and salts using qiaquick spin columns in a microcentrifuge. These are available online in convenient and compact pdf format at.