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. 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. Hispeed plasmid maxi kit email to our experts sales. Timerun, 750g yield, for ultrafast purification of up to 750g transfectiongrade plasmid. Qiagen plasmid purification protocols are based on a modified alkaline lysis. Qiagen hispeed plasmid maxi kit, 10 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 60 min. Qiagen plasmid purification handbook sierra bermeja. 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. The highspeed plasmid mini kit uses a modified alkaline lysis method and rnase treatment to obtain clear cell lysate with minimal genomic dnarna contaminants. The pureyield system requires a vacuum pump and manifold e.
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. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. These are available online in convenient and compact pdf format at. 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.
The purelink hipure plasmid maxiprep kit is designed to isolate transfectiongrade plasmid dna from e. Rna, proteins, dyes, and low molecular weight impurities are removed by a mediumsalt wash. For an overview of the qiagen web site, please turn to page 102. 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. Qiagen hispeed plasmid maxi kit, 25 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing.
Changing the direction and orientation of electric field. Plasmid or cosmid dna purification using qiagen plasmid mega and giga kits this protocol is designed for preparation of up to 2. When i use the qiagen plasmid maxi kit to prep the dna for transfection, i cannot get dna pellet. Qiagen hispeed plasmid maxi kit, 25 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 60 min. The kits feature a filter syringe for the rapid clearing.
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. Timerun, 750g yield, for ultrafast purification of up to 750g transfectiongrade. 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. For the qiagen plasmid mini kit protocol microcentrifuge 1.
Qiagen endofree plasmid maxi kit generating and harvesting plasmid dna from dh5. Hispeed plasmid maxi kit 25, 12663 from qiagen labsave. Endofree plasmid maxi kit 16 plasmid or cosmid dna purification using. Hispeed plasmid purification handbook kirschner lab. More information about biocoder can be found on my home page. For qiagen plasmid midi, maxi, mega, and giga kit protocols. The highspeed plasmid mini kit was designed for both plasmid and cosmid dna purification. I repeated three times and just can not get my dna. Plasmid purification using a qiagen hispeed plasmid maxi kit. Hispeed plasmid midi kit 25 from qiagen sample to insight. The ultrafast purification protocol, based on the remarkable selectivity of patented qiagen resin, allows the isolation of ultrapure, supercoiled plasmid dna with high. 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 maxi kit 10, 12662 from qiagen labsave. For ultrafast purification of up to 750 micro g transfectiongrade plasmid or cosmid dna.
Plasmid dna was extracted using both the highspeed plasmid kit lane 3,4 and the equivalent competitors plasmid kit lane 1,2. Please provide valid email id, we will send you account information on mail. 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. Plasmid miniprep protocol start from 25 ml overnight culture for high copy vector or 510 ml for low copy vectors volumes in brackets. Plasmid maxi 10 qiafilter plasmid maxi 10 hispeed plasmid maxi 10 na0300. 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.
Timerun, 750g yield, for ultrafast purification of up to 750g. 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. 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. Discussion plasmid purification is a basic tool of molecular biologists. Sigmas genelute hp plasmid maxiprep kits offer a simple, rapid, and costeffective method for isolating plasmid dna from recombinant e. 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. These are available online in convenient and compact pdf format.
Midi kits have a maximum binding capacity of 200 ug and the maxi 750 ug. Plasmid dna maxiprep kit product insert norgen biotek. Cas9 plasmid addgene, plasmid id 41815 as bacterial stab in agar lb agar plate containing 100. Hispeed plasmid kits contain qiafilter cartridges, hispeed tips, and qiaprecipitator. Hispeed plasmid purification handbook harvard university. Hispeed plasmid maxi kit 25 from qiagen selectscience. Qiafilterplasmidpurificationhandbook 042012 7 introduction qiafilter plasmid kits are based on the remarkable selectivity of patented qiagen resin. Please consult the material safety data sheet msds for information regarding hazards and safe handling.
What you see here is the autogenerated text ouput of the protocol that was coded up in biocoder see source code. Benchpro 2100 plasmid purification system thermo fisher. Following is the maxiprep of plasmid dna from li protocol in biocoder, a highlevel programming language for expressing biology protocols. Plasmid dna concentration plasmid dissolved in te buffer should be spectrophotometrically determined at 260 nm and.
Plasmid or cosmid dna purification using hispeed plasmid midi and maxi kits this protocol is for preparation of up to 200. 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. Endofree plasmid purification handbook 112005 11 important notes please take a few moments to read this handbook carefully before beginning the dna preparation. Qiagen plasmid purification handbook harvard university. What is your preferred maxi prep kit for plasmid dna grown. Other kit contents can be stored at room temperature 1525oc. This plasmid kit will purify plasmid dnacosmid dna from 15 ml of cultured bacterial cells using an efficient plasmid miniprep system. Mix gently by inverting the tube 46 times and incubate at room temperature for exactly 5 min. This protocol is suitable for bac dna preparation prior to transformation into sw102 strain for recombineering experiments.
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. 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. These are available online in convenient and compact pdf. 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. Plasmids and cosmids up to 50 kb in size can be purified using hispeed plasmid kit. On receipt, store the control dna and ampicillin at 20oc. Hispeed plasmid maxi kit 10 from qiagen selectscience. Each restriction digestion lanes 16 was performed in triplicate on. My experience has been with midi kit, although the maxi kit protocol is essentially identical. Plasmid dna can be purified in approximately 60 minutes, greatly reducing the time spent on purification compared to silicaresin or other membrane column methods. To help you find what youre looking for, weve gathered our plasmid kits on one page.
The system provides a rapid method for purification using a silicamembrane column. Hispeed plasmid midi and maxi kits are developed, designed, and sold for research purposes only. The kit allows for the isolation of plasmid dna with final endotoxin levels of 0. 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.
Qiagen, qiafilter, hispeed, and endofree plasmid kits, provide optimal transfection results for. By combining sdsalkaline lysis with the speed and easeofuse of silicamembrane purification, the isolate ii plasmid mini kit provides a fast method. Following the purification process, a yield of more than 900. Cross reference of genelute to qiagen nucleic acid.
Do you have any problem with qiagen plasmid maxi kit. Crisprcas9directed genome editing of cultured cells. Qiagenplasmidpurificationhandbook 042012 5 qiagen mega5 mega25 giga5 plasmid plasmidkit bufferset catalogno. Genelute hp plasmid maxiprep kit 10 12163 12263 12663 12963. What is your preferred maxi prep kit for plasmid dna grown in. Hiper plasmid dna extraction teaching kit solution based. The pureyield plasmid maxiprep system is designed to isolate highquality plasmid dna for use in eukaryotic transfection and in cellfree expression experiments.
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. For ultrafast purification of up to 750 g transfectiongrade plasmid or cosmid dna. 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. All centrifugation steps should be carried out at 16,000 x g,000 rpm. Plasmid maxi 25 qiafilter plasmid maxi 25 hispeed plasmid maxi 25 plasmid plus maxi kit 25 na0310. By eliminating hands on steps, which promote human errors, the benchpro 2100 plasmid purification system enhances experimental reproducibility and data consistency run after run. If qiagen plasmid purification kits are new to you, please visit our. Add 10 ml of buffer p3, mix gently, incubate on ice for 1520 min. Qiagen tips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column. Qiagen hispeed plasmid maxi kit, 10 preps, 150l to 250ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing. Plasmid or cosmid dna purification using qiagen hispeed. Qiagen plasmid purification handbook kirschner lab. 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.