Purelink hipure plasmid midiprep kit thermo fisher scientific. Quickstart protocol march 2016 qiagen plasmid mini, midi and maxi kits the qiagen plasmid mini kit cat. A link to the relevant protocol sheet is provided in the detailed information about the protocol. 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. You can search for protocols and protocol sheets by selecting an application, kit, or starting material. The genelute plasmid midiprep kit offers a simple, rapid, costeffective method for isolating plasmid dna from recombinant e. Qiagen plasmid purification handbook kirschner lab. Genelute hp plasmid midiprep kit has been used to isolate and purify plasmids. Purifying plasmid dna from bacterial colonies using the qiagen miniprep kit. Plasmid dna was purified from bacterial cultures using nucleobond xtra midi, nucleobond xtra maxi, and corresponding midiprep and maxiprep kits from qiagen following the protocols for each kit in each case, the maximum input volume specified by the corresponding kit protocol was processed.
For uptodate licensing information and product specific disclaimers, see the respective qiagen kit handbook or user manual. Brief considerations for plasmidcosmid purification procedures. Plasmid or cosmid dna purification using qiagen hispeed. Qiagen is the leading provider of innovative sample and assay technologies, enabling plasmid dna purification using the qiaprep spin miniprep kit. For purification of up to 100ug transfection grade plasmid or cosmid dna. Plasmid dna midiprep kits thermo fisher scientific us. All centrifugation steps should be carried out at room temperature unless otherwise noted. The kit recovers up to 20 g of high copy plasmid dna per isolation procedure. The pureyield plasmid midiprep system is designed to isolate highquality plasmid dna for use in eukaryotic transfection and in vitro expression experiments. By combining silica binding technology and the convenience of a spin column format, up to 300. Plasmid eluates are precipitated with isopropanol then passed through a prestotm plasmid concentrator using a syringe. Utilizing a modified alkaline lysis in conjunction. Monarch plasmid miniprep kit protocol monarch plasmid miniprep kits consistently produce more concentrated plasmid dna with equivalent yield, purity and functionality as compared to the leading supplier preps were performed according to recommended protocols using 1. Principles behind plasmidprinciples behind plasmid.
Qiagen plasmid purification handbook harvard university. Filter the supernatant containing the plasmid through a compress and recover filtrate in a 50 ml falcon tube. Equilibrate a qiagentip 100 by applying 4 ml buffer qbt, and allow the column to empty by gravity. 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. Endofree plasmid kits, provide optimal transfection results for ordering information, see pages 3336. Forceps are to be sterilized in fine science tools heat block at. The qiagen plasmid kits uses gravityflow qiagen anionexchange tips for efficient purification of plasmid dna. Vacuum format protocol overview for complete protocol, consult instruction manual. Qiagen qiaamp dna mini kit protocol qiagen dna qiagen dna blood mini kit protocol qiaamp dna blood midi maxi.
These are available online in convenient and compact pdf format at. Midipreps for plasmid dna purificationnucleobond xtra. Quickstart protocol march 2016 qiagen plasmid mini, midi. Find out how to transfer protocols to your qiacube. I have used mini preps on the exact same plasmid with no problem. Isolate a suitable piece of tissue and place in a uvcrosslinked 1. Formulas for qiagen kit buffers genesee scientific.
Sample preparation aurum tm plasmid midi and maxi kits. As an added convenience, the zymopure ii plasmid midiprep kit contains. The midiprep kit protocol will typically yield 100350 g plasmid dna from 1525 ml. The zymopure ii plasmid maxiprep kit features a simple spincolumn based method for the purification of up to 1.
Solutions that contain ethanol, isopropanol or mops should be sterilized by filtration only. Thermo scientific genejet plasmid maxiprep kit is qualified by isolating high copy number plasmid dna from 250 ml of overnight e. The presto plasmid dna concentration kit offers a simplified replacement for timeconsuming plasmid dna precipitation procedures. For the qiagen plasmid mini kit protocol microcentrifuge 1. Genelute plasmid dna midiprep kit 35 purifications gen. Qiagen plasmid purification protocols are optimized for use with cultures grown in standard luria bertani. Plasmid purification using a qiagen hispeed plasmid maxi kit.
Zymopure plasmid isolation kits provide the fastest and simplest method available to efficiently isolate transfection quality plasmid dna from e. Hispeed plasmid midi and maxi kit for rapid purification of transfection grade. Genelute hp plasmid dna midiprep kit 25 purifications. Features and benefits from harvested bacterial culture to pure plasmid dna in 30 minutes or less up to 350. The pureyield plasmid midiprep systema is designed to isolate highquality plasmid dna for use in eukaryotic transfection and in vitro expression experiments. For larger constructs qiagen plasmid kits or the largeconstruct kit are recommended. Thermo scientific genejet plasmid miniprep kit utilizes an exclusive silicabased membrane technology in the form of a convenient spin column. Qiagen plasmid purification handbook 082003 9 product use limitations qiagen, qiafilter, and endofree plasmid kits are developed, designed and sold for research purposes only. Highlights efficienthigh yields of up to 20 g of high quality plasmid dna fastpr. Plasmids and cosmids up to 50 kb in size can be purified using hispeed plasmid kit. Thermo scientific genejet plasmid midiprep kit is qualified by isolating high copy plasmid dna from 50 ml of overnight e. Here we introduce the miraprep, a rapid protocol that allows isolation of plasmid dna using commercial miniprep kits. Plasmid midiprep preparation is a basic technique performed in most molecular biology laboratories.
Qiagen plasmid plus midi kit, 25 preps, 25 to 50ml culture volume. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. Plasmid or cosmid dna purification using qiagen plasmid midi and maxi kits. Multi sample processing is often required to complete both plasmid isolation and subsequent downstream experimentation. Qiagen high speed midiprep protocol qiagen high speed midiprep protocol image quotes, qiagen high speed midiprep protocol quotes and saying, inspiring quote pictures, quote pictures.
Invitrogen plasmid midiprep kits offer a range options to eliminate the challenges raised by this level of workflow processing. The qiagen largeconstruct kit is designed for the purification of. Hispeed plasmid purification handbook kirschner lab. I have done lots of mini preps before but this is my first midi prep. Plasmid or cosmid dna purification, advanced silica technology, manual. Qiagen plasmid plus midi kit 25 from qiagen sample to insight. Vigorously shake the buffer bottle before use to ensure lyseblue particles are. 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. The qiaprep spin miniprep kit is designed for isolation of up to 20. For uptodate licensing information and productspecific disclaimers, see the respective qiagen kit handbook or user manual.
Qiagen plasmid purification handbook sierra bermeja. The system provides a rapid method for purification using a silicamembrane column. Plasmid purification is a basic tool of molecular biologists. The kit passes qc requirements if the purified plasmid dna has an a 260 a. Hispeed plasmid midi and maxi kits protocol before staring make sure that rnase a has been added to p1 buffer. Qiagen plasmid purification handbook home facebook. Purifying plasmid dna using the qiagen miniprep kit protocol. Plasmid or cosmid dna purification using qiagen plasmid mega and giga kits this protocol is designed for preparation of up to 2. Principles behind plasmidprinciples behind plasmid purifications. Start a 2 ml for mengo clones use starter culture in the morning from an. Hispeed plasmid purification handbook harvard university. Although the development of plasmid isolation kits utilizing silica spin columns reduced the time and labor spent on plasmid purification, achieving large plasmid dna yields still requires significant time and effort. Print bookmark share pdf 1126kb english format file size language download get adobe reader contact qiagen.