Qiagen plasmid purification handbook harvard university. Timerun, up to 1mg yield, ideal transfection of most cell lines, cloning. Qiagen plasmid midi kit vs zymopure plasmid midiprep kit 25 preps this page will help you compare kit specs and protocols across manufacturers. The endotoxin content of the purified plasmid dna was determined using the kineticqcl kinetic chromogenic assay. Mini 25 12123 mini 100 12125 midi 25 12143 midi 100 12145 maxi 10 12162. Sigmas genelute hp plasmid maxiprep kits offer a simple, rapid, and costeffective method for isolating plasmid dna from recombinant e. I repeated three times and just can not get my dna. Mar 01, 2019 full plasmid dna extraction completed using a qiagen maxiprep kit. Do you have any problem with qiagen plasmid maxi kit. Please consult the material safety data sheet msds for information regarding hazards and safe handling. The midiprep kit protocol will typically yield 100350 g plasmid dna from 1525 ml bacterial culture, at a purity that is comparable to that achieved by two passes through cesium chloride gradients. 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. The genopure plasmid midi kit prepares highly purified plasmid dna in medium quantities using a modified alkaline lysis method.
For the fastest, most convenient purification of up to 10 mg transfectiongrade plasmid dna. Manual centrifugation manual centrifugation manual. Qiagen plasmid maxi kit 10, 12162 from qiagen labsave. If you want to perform plasmid purification without using a kit, you can find a protocol for kit free plasmid miniprep at the bottom of this page. For qiagen plasmid midi, maxi, mega, and giga kit protocols centrifugation tubes or vessels with suitable capacity for the volumes specified in the. The kits feature a filter syringe for the rapid clearing. Qiagen circulating nucleic acid kit suggest leaving columns for 23 min after adding ave buffer at the elution step. Highquality plasmid dna is eluted in a small volume of tris buffer or deionized water. Filmed at the university of northampton, by adam gregory. The kit utilizes silicabased membrane technology in the form of convenient spin column. The purelink hipure plasmid midiprep kit is designed to isolate transfectiongrade plasmid dna from e.
When i use the qiagen plasmid maxi kit to prep the dna for transfection, i cannot get dna pellet. Qiafilter plasmid midi kit 25 from qiagen selectscience. Quickstart protocol march 2016 qiagen plasmid mini, midi. Hispeed plasmid midi kit 25 from qiagen sample to insight. Qiaquick gel extraction kit protocol using a microcentrifuge. 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. 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. The pureyield plasmid midiprep system is designed to purify 100200g of plasmid dna with a 260 a 280 1. Qiagen plasmid plus maxi kit vs zymopure plasmid maxiprep kit 25 preps this page will help you compare kit specs and protocols across manufacturers. With the rneasy midi kit, total rna is easily purified from 5 x 10 6 to 1 x 10 8 animal or human cells, 20250 mg animal or human tissues, and 2 x 10 7 to 5 x 10 8 yeast cells. Qiagen tips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column.
Print bookmark share pdf 460kb english format file size language download. Pureyield plasmid maxiprep system instructions for use of products a2391, a2392 and a2393. Qiagen mini mini midi midi maxi maxi maxi plasmidkit 25 100 25 100 10 25 100. My experience has been with midi kit, although the maxi kit protocol is essentially identical. This kit can also be used for dna cleanup from enzymatic reactions see page 8. Up to 400 mg agarose can be processed per spin column. For qiagen buffer compositions, please see the qiagen buffers page. Qiagen hispeed plasmid midi kit, 25 preps, 50l to 150ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 25 preps this page will help you compare kit specs and protocols across manufacturers. Qiagen plasmid plus maxi kit 25, 12963 from qiagen labsave. For purification of up to 10 mg transfectiongrade plasmid or cosmid dna. Oct 11, 2005 the qiagen hispeed plasmid kit comes in midi and maxi formats. Hispeed plasmid purification handbook harvard university. Gently mix by flicking the bottom of the tube with your finger a few times. Qiaamp dna mini kit and qiaamp dna blood mini kit handbook.
If you want to perform plasmid purification without using a kit, you can find a protocol for kitfree plasmid miniprep at the bottom of this page. Mix gently by inverting the tube 46 times and incubate at room temperature for exactly 5 min. The dna isolated by this kit is ready for downstream applications such as restriction enzyme digestion, sequencing, pcrrealtime pcr and other downstream. Qiagen plasmid plus maxi kit, 25 preps, 50 to 200ml culture volume, transfectiongrade plasmid or cosmid dna purification, advanced silica technology, manual processing, 20 min. Small scale purification miniprep from less than 5 ml of bacterial culture is a quick way for clone verification or dna isolation, followed by further enzymatic reactions polymerase chain reaction and restriction enzyme digestion. Filmed using an iphone xsmax and inbuilt imovie features. Qiagentips can be left unattended, since the flow of buffer will stop when the meniscus reaches the upper frit in the column. Na0400 genelute hp endotoxinfree plasmid maxiprep kit 10 preps. Timerun, up to 1mg yield, ideal transfection of most cell lines, cloning, enzymatic. L of competent cells in a microcentrifuge or falcon tube. 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. Lyophilized qiagen protease can be stored at room temperature 1525c for up to. Qiagen hispeed midi plasmid dna purification kit biocompare.
Quickstart protocol march 2016 qiagen plasmid plus midi kit the qiagen plasmid plus midi kit cat. Purify up to 20 samples 10 minutes handsontime75 minutes overall. The dna isolated by this kit is ready for downstream applications such as restriction enzyme digestion, sequencing, pcrrealtime pcr and other downstream experiments. Qiagen plasmid plus midi kit, 25 preps, 25 to 50ml culture volume, transfectiongrade plasmid or cosmid dna purification, advanced silica technology, manual processing, 20 min. The genelute hp plasmid miniprep kit offers a simple, rapid, and costeffective method for isolating plasmid dna from recombinant e. Recovering plasmid dna from bacterial culture addgene. Click on the product or protocol links to be directed to the manufacturers pages. Plasmid size the protocols are suitable for purifying most plasmids ranging from. Quickstart protocol march 2016 qiagen plasmid plus midi kit. Use the purelink quick plasmid miniprep kit to isolate high quality plasmid dna up to 30 g from e. This protocol is designed for purification of up to 40. Plasmid dna was purified from 25 ml and 100 ml cultures of the indicated e. Qiagen plasmid purification protocols are optimized for use with cultures grown in standard luria bertani lb medium to a cell density of approximately 34 x 109.
Qiagen plasmid purification handbook 082003 9 product use limitations qiagen, qiafilter, and endofree plasmid kits are developed, designed and sold for research purposes only. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. For ultrafast purification of up to 750 g transfectiongrade plasmid or cosmid dna. 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 hispeed plasmid midi kit, 25 preps, 50l to 150ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, plasmid dna purification from e. Quickclean ii plasmid miniprep kit provides a fast, simple, and costeffective method for purification of up to 20. Sureprep plasmid purification kits contain sureprep columns of different sizes mini, midi, maxi, mega and giga, appropriate buffers and rnase a. Timerun, up to 1mg yield, for the fastest, most convenient purification of up to 1mg.
Qiagen plasmid plus maxi kit vs zymopure plasmid maxiprep. 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. Qiagen buffer also works for epoch life sciences spin columnswhich are sold in bulk at a much lower price. Do you have a protocol for the isolation of bac dna using the qiagen plasmid midi kit. Thermo scientific genejet plasmid midiprep kit, sufficient for 100 preps, is designed for large scale isolation of high quality plasmid dna from recombinant e. All others centrifugation steps should be carried out at room. If you will be using a kit, follow the kits instructions. Ethanol is added to the lysate to provide ideal binding conditions. Protip transformation efficiencies will be approximately 10fold lower for ligation of inserts to vectors than for an intact control plasmid. Quickstart protocol march 2016 qiagen plasmid mini, midi and maxi kits the qiagen plasmid mini kit cat. The geneaid midi plasmid kit uses prepacked anionexchange resin columns to purify plasmid dna from 50300 ml of cultured bacterial cells. Qiagen plasmid maxi kit, 10 preps, 100 to 500ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, 500g yield, 160 min. Plasmid dna prepared by tianprep mini plasmid kit is suitable for. The qiagen plasmid kits uses gravityflow qiagen anionexchange tips for efficient purification of plasmid dna.
Nov 18, 2015 manual centrifugation manual centrifugation manual. 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. For purifying plasmid dna from escherichia coli cells, the qiagen spin miniprep kit produces quite reliable results. For fast purification of up to 10 mg transfectiongrade plasmid or cosmid dna. Full qiagen maxiprep plasmid dna extraction with steps. Description quickclean ii plasmid miniprep kit provides a fast, simple, and costeffective method for purification of up to 20. If you will be using a kit, follow the kit s instructions.
The qiagen hispeed plasmid kit comes in midi and maxi formats. Qiagen plasmid midi kit vs zymopure plasmid midiprep kit 25. May 09, 2012 for the love of physics walter lewin may 16, 2011 duration. For the love of physics walter lewin may 16, 2011 duration. Plasmid or cosmid dna purification using qiagen plasmid midi and maxi kits. The protocol below is meant to describe the general procedure for purifying plasmid dna from bacterial cultures. Sureprep columns are polypropylene columns containing sureprep anionexchange silica resin packed between two inert filter elements which can.
Equilibrate a qiagentip 100 by applying 4 ml buffer qbt, and allow the column to empty by gravity flow. Qiagen plasmid plus midi kit 25 from qiagen selectscience. Quick start protocol 1 1 1 1 provided in a 10 mgml or 100 mgml solution. Timerun, up to 1mg yield, for the fastest, most convenient purification of up to. Midi kits have a maximum binding capacity of 200 ug and the maxi 750 ug. If qiagen plasmid purification kits are new to you, please visit our. Equilibrate a qiagen tip 100 by applying 4 ml buffer qbt, and allow the column to empty by gravity flow. Add 10 ml of buffer p3, mix gently, incubate on ice for 1520 min. Plasmids and cosmids up to 50 kb in size can be purified using hispeed plasmid kit.
Rna, proteins, dyes, and low molecular weight impurities are removed by a mediumsalt wash. Maxi plasmid dna kit pm010, pm025 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. Timerun, 100g yield, ideal for transfection, cloning, sequencing, lysate clearing without. 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.