Cross reference of genelute to qiagen nucleic acid. Plasmid purification using a qiagen hispeed plasmid maxi kit. This plasmid kit will purify plasmid dnacosmid dna from 15 ml of cultured bacterial cells using an efficient plasmid miniprep system. By eliminating hands on steps, which promote human errors, the benchpro 2100 plasmid purification system enhances experimental reproducibility and data consistency run after run. Plasmid or cosmid dna purification using qiagen plasmid mega and giga kits this protocol is designed for preparation of up to 2. Qiagen hispeed plasmid maxi kit, 25 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 60 min.
Genelute hp plasmid maxiprep kit 10 12163 12263 12663 12963. Hiper plasmid dna extraction teaching kit solution based. The ultrafast purification protocol, based on the remarkable selectivity of patented qiagen resin, allows the. Method outline this method is modified from the qiagen plasmid purification handbook protocol for plasmid or cosmid dna purification using the endofree plasmid maxi kit pages 3640. Rna, proteins, dyes, and low molecular weight impurities are removed by a mediumsalt wash. Qiagen plasmid purification handbook harvard university. Changing the direction and orientation of electric field. What is your preferred maxi prep kit for plasmid dna grown. The purified dna is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95100% accuracy, bacterial. Hispeed plasmid midi and maxi kits are developed, designed, and sold for research purposes only. The purelink hipure plasmid maxiprep kit is designed to isolate transfectiongrade plasmid dna from e.
Hispeed plasmid maxi kit 10, 12662 from qiagen labsave. Hispeed plasmid kits contain qiafilter cartridges, hispeed tips, and qiaprecipitator. Each restriction digestion lanes 16 was performed in triplicate on. Qiagen hispeed plasmid midi kit, 25 preps, 50l to 150ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, oct 11, 2005 the qiagen hispeed plasmid kit comes in midi and maxi formats. Hispeed plasmid maxi kit 25, 12663 from qiagen labsave. Qiagens plasmid plus maxi kit is frequently used by me because its cost effective and not time consuming with high yield of dna as well as it uses a vacuum manifold. The kit is stable for 12 months from the date of manufacture store control dna and ampicillin at 20oc store 6x gel loading buffer, li cells with plasmid and resuspension solution at 28oc other kit contents can be stored at room temperature 1525oc hiper plasmid dna extraction teaching kit solution based.
Qiafilterplasmidpurificationhandbook 042012 7 introduction qiafilter plasmid kits are based on the remarkable selectivity of patented qiagen resin. Discussion plasmid purification is a basic tool of molecular biologists. Cas9 plasmid addgene, plasmid id 41815 as bacterial stab in agar lb agar plate containing 100. For larger constructs qiagen plasmid kits or the largeconstruct kit are recommended plasmidcosmid copy number plasmid and cosmids vary in copy number, depending on the origin of replication they contain, their size, and the size of insert. Addgenes repository contains over 80,000 plasmids, which includes over 90 kits. The kit is able to purify plasmids up to,000 bp in size, and typical yields from a 100 ml culture for a high copy number plasmid are between 0. Plasmid dna was extracted using both the highspeed plasmid kit lane 3,4 and the equivalent competitors plasmid kit lane 1,2.
Hispeed plasmid midi kit 25 from qiagen sample to insight. To help you find what youre looking for, weve gathered our plasmid kits on one page. The geneaid maxiprep plasmid kit was designed for plasmid dna purification using prepacked anionexchange resin maxiprep plasmid columns to purify plasmid dna from 2001600 ml of cultured bacterial cells. Qiagen plasmid purification handbook 082003 5 kit contents qiagen plasmid kits qiagen plasmid kit midi 25 midi 100 maxi 10 maxi 25 maxi 100 catalog no. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. Hispeed plasmid maxi kit email to our experts sales. My experience has been with midi kit, although the maxi kit protocol is essentially identical. Plasmid maxi 10 qiafilter plasmid maxi 10 hispeed plasmid maxi 10 na0300. Following the purification process, a yield of more than 900. For ultrafast purification of up to 750 g transfectiongrade plasmid or cosmid dna.
For ultrafast purification of up to 750 micro g transfectiongrade plasmid or cosmid dna. What is your preferred maxi prep kit for plasmid dna grown in. I repeated three times and just can not get my dna. Mix gently by inverting the tube 46 times and incubate at room temperature for exactly 5 min. Benchpro 2100 plasmid purification system thermo fisher.
Plasmid or cosmid dna purification using qiagen hispeed plasmid midi kits this protocol is for preparation of up to 200g of high or lowcopy plasmid or cosmid dna using the qiagen hispeed plasmid midi kit. Plasmids and cosmids up to 50 kb in size can be purified using hispeed plasmid kit. As research moves from dna sequencing to analysis of gene function, the need has increased for rapid methods for isolating large quantities of highquality plasmid dna. Qiagen endofree plasmid maxi kit generating and harvesting plasmid dna from dh5.
Plasmid maxi 25 qiafilter plasmid maxi 25 hispeed plasmid maxi 25 plasmid plus maxi kit 25 na0310. Hispeed plasmid purification handbook kirschner lab. Qiagenplasmidpurificationhandbook 042012 5 qiagen mega5 mega25 giga5 plasmid plasmidkit bufferset catalogno. Qiagen hispeed plasmid maxi kit, 10 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 60 min. Please provide valid email id, we will send you account information on mail. Sigmas genelute hp plasmid maxiprep kits offer a simple, rapid, and costeffective method for isolating plasmid dna from recombinant e. The highspeed plasmid mini kit uses a modified alkaline lysis method and rnase treatment to obtain clear cell lysate with minimal genomic dnarna contaminants. More information about biocoder can be found on my home page. The kit allows for the isolation of plasmid dna with final endotoxin levels of 0. Plasmid dna maxiprep kit product insert norgen biotek.
The system provides a rapid method for purification using a silicamembrane column. Plasmid miniprep protocol start from 25 ml overnight culture for high copy vector or 510 ml for low copy vectors volumes in brackets. For an overview of the qiagen web site, please turn to page 102. The highspeed plasmid mini kit was designed for both plasmid and cosmid dna purification. Qiagen plasmid purification protocols are based on a modified alkaline lysis. Hispeed plasmid purification handbook harvard university. These are available online in convenient and compact pdf format. Hispeed plasmid maxi kit 25 from qiagen selectscience.
Hispeed plasmid maxi kit 10 from qiagen selectscience. For the qiagen plasmid mini kit protocol microcentrifuge 1. Qiagen plasmid purification handbook sierra bermeja. Timerun, 750g yield, for ultrafast purification of up to 750g. Timerun, 750g yield, for ultrafast purification of up to 750g transfectiongrade. By combining sdsalkaline lysis with the speed and easeofuse of silicamembrane purification, the isolate ii plasmid mini kit provides a fast method. Plasmid or cosmid dna purification using hispeed plasmid midi and maxi kits this protocol is for preparation of up to 200. Qiagen hispeed plasmid maxi kit, 25 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing. These are available online in convenient and compact pdf. Qiagen plasmid purification handbook kirschner lab. What you see here is the autogenerated text ouput of the protocol that was coded up in biocoder see source code.
Do you have any problem with qiagen plasmid maxi kit. Plasmid dna concentration plasmid dissolved in te buffer should be spectrophotometrically determined at 260 nm and. If you have questions about any of the information in the bench guide, or about molecular biology in general, please call your local qiagen technical service department or your local distributor see back cover. Midi kits have a maximum binding capacity of 200 ug and the maxi 750 ug. Qiagentips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column. The ultrafast purification protocol, based on the remarkable selectivity of patented qiagen resin, allows the isolation of ultrapure, supercoiled plasmid dna with high. All centrifugation steps should be carried out at 16,000 x g,000 rpm. The pureyield system requires a vacuum pump and manifold e. Following is the maxiprep of plasmid dna from li protocol in biocoder, a highlevel programming language for expressing biology protocols. The protocol for qiafilter plasmid kits is optimized for use with cultures grown in standard luria bertani lb medium see page 36, grown to a cell density of. This protocol is suitable for bac dna preparation prior to transformation into sw102 strain for recombineering experiments. The kits feature a filter syringe for the rapid clearing. Trueblue lysis buffer an optional color indicator is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization. For qiagen plasmid midi, maxi, mega, and giga kit protocols.
In accordance with qiagens isocertified quality management system, each lot of the rneasy mini kit, rneasy protect mini kit, and rneasy plant mini kit is tested against predetermined specifications to ensure consistent product quality. Endofree plasmid purification handbook 112005 11 important notes please take a few moments to read this handbook carefully before beginning the dna preparation. Add 10 ml of buffer p3, mix gently, incubate on ice for 1520 min. Qiagen, qiafilter, hispeed, and endofree plasmid kits, provide optimal transfection results for. Qiagen s plasmid plus maxi kit is frequently used by me because its cost effective and not time consuming with high yield of dna as well as it uses a vacuum manifold. Crisprcas9directed genome editing of cultured cells. Plasmid or cosmid dna purification using qiagen hispeed. In the 3rd time i retransformed my plasmid and pick a single colony and culture in 25 ml lb medium containing ampicilian, incubating for approximately 6 h at 37c with vigorous shaking and. Please consult the material safety data sheet msds for information regarding hazards and safe handling.
Qiagen tips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column. Other kit contents can be stored at room temperature 1525oc. These are available online in convenient and compact pdf format at. Plasmid dna can be purified in approximately 60 minutes, greatly reducing the time spent on purification compared to silicaresin or other membrane column methods.
If qiagen plasmid purification kits are new to you, please visit our. Qiagen hispeed plasmid midi kit, 25 preps, 50l to 150ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 25 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 60 min. When i use the qiagen plasmid maxi kit to prep the dna for transfection, i cannot get dna pellet. The pureyield plasmid maxiprep system is designed to isolate highquality plasmid dna for use in eukaryotic transfection and in cellfree expression experiments. The maxiprep kit protocol will typically yield 500850 g plasmid dna from 100200 ml bacterial culture, at a purity that is comparable to that achieved by two passes through cesium chloride gradients. On receipt, store the control dna and ampicillin at 20oc. Qiagen hispeed plasmid maxi kit, 10 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing.
The isolate ii plasmid mini kit provides a simple, efficient columnbased method for the isolation of plasmid dna from bacterial cultures, without the need for hazardous reagents such as phenol. For larger constructs qiagen plasmid kits or the largeconstruct kit are recommended plasmid cosmid copy number plasmid and cosmids vary in copy number, depending on the origin of replication they contain, their size, and the size of insert. Endofree plasmid maxi kit 16 plasmid or cosmid dna purification using. Timerun, 750g yield, for ultrafast purification of up to 750g transfectiongrade plasmid.
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