The qiacube is preinstalled with protocols for purification of plasmid dna, genomic dna, rna, viral nucleic acids, and proteins, plus dna and rna cleanup. The purchaser must determine the suitability of the product for its particular use. Typical yields from low copy number plasmids are highly dependent upon culture conditions and vectorhost strain combinations. These plasmid kits feature zymopure ii technology, including. Techniques in molecular biology methods for plasmid dna isolation 3 other notes on this plasmid miniprep technique once cells have been lysed, mixing should be done thoroughly but gently, to avoid breaking plasmid and bacterial chromosomal dna.
Zymopure ii plasmid kits are the fastest and simplest plasmid kits available to efficiently isolate transfectiongrade plasmid dna from e. Purifying plasmid dna using the qiagen miniprep kit protocol. The procedure can be performed in 40 minutes compactprep mega or 50 minutes compactprep giga and uses a vacuum pump and centrifuge. Thomson instrument company is not affiliated with millipore, emd, qiagen, bio101 or their products step 1 step 2 miniprep determine dna concentration mgsl mega column. Quick start procedure this quick start procedure is for. Add the provided rnase a solution to buffer mx1, mix and store at 4 c. This new plasmid dna isolation protocol will significantly reduce time and labor without increasing costs. The pureyield plasmid midiprep system is designed to isolate highquality plasmid dna for use in eukaryotic transfection and in vitro expression experiments. Genelute mammalian genomic dna miniprep 10 genelute mammalian genomic dna miniprep 70 genelute blood genomic dna kit 70 60506. Do not vortex after cell resuspension, but mix by inversion. For uptodate licensing information and product specific disclaimers, see the respective qiagen kit handbook or user manual. Qiagen plasmid purification handbook kirschner lab. Dna purification and analysis maximize sample yield, purity and integrity. Rneasy kits and rneasy protect kits are available in midi and maxi formats.
Formulas for qiagen kit buffers for long term storage, all buffers should be sterilized by filtration or autoclaving. The blue and purple qiagen columns are identical in formulation. Dna isolation with qiagen genomictips this protocol applies to. The endofree maxiprep kit from qiagen or the genejet maxiprep kit from thermo fisher were used for maxi preparations. Throughout the remainder of this document the supplied cell resuspension solution cra, cell lysis solution cla and neutralization solution nsb are referred to as cell resuspension solution, cell lysis. After addition of ethanol, the sample is loaded onto an rneasy mini spin. All remaining components should be stored at room temperature. Rich media are not recommended for plasmid preparation with qiagentips. Solutions that contain ethanol, isopropanol or mops should be sterilized by filtration only. A protocol for isolation of plasmid dna for use in eukaryotic transfection and in vitro expression experiments. Mega, giga purelink hipure filter maxi plasmid dna.
Grow 25100 ml of bacteria culture for one midi prep and 100500 ml for maxi prep. Qiagentips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column. For the qiagen plasmid mini kit protocol microcentrifuge 1. Plasmid or cosmid dna purification using qiagen plasmid mega and giga kits this protocol is designed for preparation of up to 2. Qiagen endofree kit purelink expi kit qiagen protocol, troubleshooting and other methodology information contact experts in qiagen to get answers. Purified dna usually is good enough for transfection and microinjection. Qiagen plasmid mega and giga kits en print bookmark share pdf 299kb english format file size language download get adobe reader contact qiagen. Genomic dna prep protocol dasen lab genomic dna prep for pcr and southern 1. Hispeed plasmid purification handbook harvard university. Should any product fail to perform satisfactorily due to any reason other.
Alternatively, qiagentip 20 may be placed in qiarack 1 figure 1b. For routine giga preparation of lowcopy plasmids, additional buffers p1. Qiagen plasmid purification handbook harvard university. They can be stored for at least 2 years without showing any reduction in performance, capacity, or quality of separation. These are available online in convenient and compact pdf. What you see here is the autogenerated text ouput of the protocol that was coded up in biocoder see source code. 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. 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. The range of protocols available is continually expanding, and additional qiagen protocols can. Qiagen plasmid purification handbook sierra bermeja. Compactprep plasmid megagiga purification handbook qiagen. They are interchangeable and can be reused multiple times after treatment with hydrochloric acid, reequilibration, and wash.
Store solution irnase a at 28c after being combined see page 7. Preparation and lysis of bacterial cell cultures note. Purifying plasmid dna from bacterial colonies using the qiagen miniprep kit. I want to use a endofree giga prep kit from qiagene. Very likely this protocol can be used with other similar columns. Buffer c1 contains chaotropic salts, which may form reactive compounds when combines with bleach. Up to 10 mg highcopy plasmid dna is purified from 2. Protocol buffer c1 contains acetic acid, wear gloves or protective eyewear when handling.
Qiagen plasmid midi prep can be used to purify up to 100 mg of plasmid or cosmid dna. 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. Qiaquick gel extraction kit protocol using a microcentrifuge. Invitrogen purelink quick plasmid miniprep kit was used in tacloning in order to qiagen plasmid mini prep kit was used to perform plasmid preparation. Here, we videorecorded the general procedures of miniprep through the qiagen s qiaprep 8 miniprep kit, aiming to introducing this highly efficient technique to the general beginners for molecular biology techniques. Qiagen plasmid purification protocols are optimized for use with cultures grown in standard luria bertani lb medium to a cell density of approximately 3 4 x 910 cellsml, which typically corresponds to a pellet wet weight of approximately 3 gliter medium. Qiagen maxi prep kit low endotoxin without additional steps. The qiagen plasmid giga kit uses gravityflow qiagen tip 0 anionexchange tips for efficient purification of plasmid dna. Following is the miniprepqiagen kit protocol in biocoder, a highlevel programming language for expressing biology protocols. Qiagen plasmid maxi prep can be used to purify up to 500 mg of plasmid or cosmid dna. The qiaprep spin miniprep kit is designed for isolation of up to 20. Qiaprep miniprep handbook washington state university. More information about biocoder can be found on my home page.
Qiagen qiaprep mini prep dna purification method cn 27014 plasmid isolation procedure taken directly from the qiagen handbook with additional notation for clarity 1 resuspend the bacterial pellet in 250 l of buffer p1. Plasmid or cosmid dna purification using qiagen plasmid mega and giga kits. Add 24 ml 50 preps or 120 ml 250 preps 98100% ethanol to wn buffer before use. These are available online in convenient and compact pdf format. For purification of up to 10 mg transfectiongrade plasmid or cosmid dna. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. Use a high copynumber plasmid to obtain a good yield of plasmid dna. The purelink hipure expi plasmid gigaprep kit enables isolation of the highest levels of transfectiongrade plasmid dna from a single li giga prep using an enhanced anion exchange resin.
Qiagentips may be placed in tubes using tip holders provided with the kits figure 1a. Setup of qiagentips awith tip holder or bwith qiarack 1. Review and cite plasmid dna isolation protocol, troubleshooting and other methodology information contact experts in plasmid dna isolation to get answers. These are available online in convenient and compact pdf format at. The new qiagen plasmid prep service provides ultrapure plasmid dna in milligram to. Do not add bleach or acidic solutions directly to the preparation waste.
Qiagen plasmid purification handbook 082003 7 endofree plasmid kits endofree plasmid kit maxi 10 mega 5 giga 5 catalog no. Add 40 ml 50 preps or 180 ml 250 preps 98100% ethanol to buffer ws before use. For qiagen plasmid midi, maxi, mega, and giga kit protocols. A resource portal for protocols in bioscience including molecular biology, cell biology, immunology, plant biology, developmental biology, neuroscience, bioinformatics, genetics. Qiagen plasmid mega and giga kits en print bookmark share pdf 299kb english. The purelink hipure plasmid gigaprep kit also includes vacuum filtration columns that provide bacterial filtration without centrifugation, enabling. The qiafilter megagiga cartridge is designed for use with a 1 liter, 45mmneck glass bottle e. Provides protocol of isolating doublestranded dna from plasmid or cosmid by the alkaline lysis procedure followed by equilibrium ultracentrifugation in cesium chlorideethidium bromide gradients and. Unused columns can be cheaply purchased in bulk from epoch biolabs. Qiagen tips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column. Endofree plasmid dna giga kit components are guaranteed for at least 12 months from the date of purchase when stored as follows. Qiafilterplasmidpurificationhandbook 042012 7 introduction qiafilter plasmid kits are based on the remarkable selectivity of patented qiagen resin.
Instructions for use of products a2492, a2495, a2496. If you have questions about product specifications or performance, please call qiagen technical services or your local distributor see back cover or visit. Formulas for qiagen kit buffers genesee scientific. Purelink hipure plasmid dna megaprep and gigaprep kits. The ultrafast purification protocol, based on the remarkable selectivity of patented qiagen resin, allows the isolation of ultrapure, supercoiled plasmid dna. The plasmid preparation procedure is performed as usual. The purelink hipure plasmid gigaprep kit also includes vacuum filtration columns that provide bacterial filtr. Up to 400 mg agarose can be processed per spin column. Culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium. Emc skip over night culture go directly from plate into 100 mls of lb. No cell clumps should be visible after resuspension of the pellet.