Harvest the cells by centrifugation at 6000 x g 6,000 rpm in a ja20 rotor for 5 min. Purifying plasmid dna using the qiagen miniprep kit protocol. Qiagen qiafilter plasmid midi kit, 25 preps, 25 to 50ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, 110 min. Qiaprep miniprep handbook washington state university. Qiagen plasmid plus maxi kit vs zymopure plasmid maxiprep. Qiafilter plasmid midi kit 25 from qiagen selectscience. 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. Hispeed plasmid purification handbook harvard university. Qiagen plasmid maxi kit 25 from qiagen sample to insight. Click on the product or protocol links to be directed to the manufacturers pages. The procedure can be performed in 20 midi and maxi, 40 mega, or 50 minutes. For purification of up to 10 mg transfectiongrade plasmid or cosmid dna.
You will purify a plasmid construct containing a gene fusion product between cus f and yellow fluorescent protein cusfyfp using. Qiagen plasmid purification handbook sierra bermeja. Formalinfixed, paraffin embedded ffpe tissue remains the standard of genomic dna was purified using the qiaamp dna ffpe tissue kit qiagen. Plasmid or cosmid dna purification using qiagen plasmid mini kit.
Inoculate 1 ml of the starter culture in 100 ml of lb medium plus ampicillin and grow over night. Plasmid dna extraction using the presto midi plasmid kit from geneaid. May 09, 2012 plasmid dna extraction using the presto midi plasmid kit from geneaid. Qiagen buffer also works for epoch life sciences spin columnswhich are sold in bulk at a much lower price. Buffer compositions are given per liter of solution. Compactprep midi and maxi kits qiagen plasmid plus midi, maxi, mega and giga kits 96well. Both the qiaprep spin miniprep kits and qiagen plasmid plus kits work with the manifold, which can handle up to 24 samples at the same time.
Do you have any problem with qiagen plasmid maxi kit jan292006 hi dear all, when i use the qiagen plasmid maxi kit to prep the dna for transfection, i cannot get dna pellet. Timerun, up to 1mg yield, ideal transfection of most cell lines, cloning, enzymatic. Optimized buffers and reagents and triedandtested exposure times and column. Qiagen plasmid plus midi kit 25 from qiagen selectscience. Qiagen plasmid purification handbook kirschner lab. Low elution volumes yield highly concentrated plasmid dna for direct use without ethanol precipitation. Plasmid or cosmid dna purification using qiagen plasmid midi and maxi kits this protocol is designed for preparation of up to 100 g of high or lowcopy plasmid or cosmid dna using the qiagen plasmid midi kit, or up to 500 g using the qiagen plasmid maxi kit. Qiagen plasmid purification handbook gel electrophoresis. Qiagen plasmid prep buffer compositon and preparation. Qiagen plasmid maxi kit, 100 preps, 100 to 500ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, 500g yield, 160 min.
Buffer calculations are base on tris base adjusted to ph with hcl triscl. Rneasy mini handbook rneasy mini kit for purification of total rna from animal cells, animal tissues, bacteria, and yeast, and for rna cleanup rneasy protect mini kit for immediate stabilization of rna in harvested animal tissues and subsequent total rna purification rneasy plant mini kit for purification of total rna from plants and filamentous fungi fourth edition june. Qiagen plasmid plus kits provide a novel patentpending method for extremely fast and easy largescale preparation of transfectongrade plasmid dna. For qiagen buffer compositions, please see the qiagen buffers page. The innovative binding technology and optimized protocol enable elution. Do you have any problem with qiagen plasmid maxi kit. Qiagen plasmid plus midi kit 25 from qiagen sample to.
Please consult the material safety data sheet msds for information regarding hazards and safe handling. Do not autoclave isopropanolcontaining buffer, but sterilize by filtration instead. Plasmid dna purification using qiagen plasmid plus midi kits 18. Qiagen originals pure plasmids, genuine kits learn more. If properly stored, all kit components are stable until the expiration date printed on the label. For fast purification of up to 10 mg transfectiongrade plasmid or cosmid dna. Qiagen plasmid maxi kit 100, 12165 from qiagen labsave. Qiagen clinical kits compatible with the qiavac 24 plus application kit accessories genomic and viral dna purification from blood and body fluids qiaamp dna blood kits vacconnectors and vacvalves qiaamp dna mini kit vacconnectors and vacvalves viral rna purification from plasma. Promega wizard kit was a long complicated kit, i would choose the. The geneaid midi plasmid kit uses prepacked anionexchange resin columns to purify plasmid dna from 50300 ml of cultured bacterial cells. The purchaser must determine the suitability of the product for its particular use.
The results illustrate the very high binding capacity of the columns in the isolate ii plasmid mini kit. Qiagen plasmid plus maxi kit vs zymopure plasmid maxiprep kit 25 preps this page will help you compare kit specs and protocols across manufacturers. Hispeed plasmid midi and maxi kits protocol before staring make sure that rnase a has been added to p1 buffer. Qiagen plasmid maxi kit, 10 preps, 100 to 500ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, 500g yield, 160 min. Qiagen plasmid plus kits for the fastest and most convenient purification. Comparison of plasmid dna yields using different methods qiagen plasmid plus highyield protocol qpp. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. In this protocol we show how to use smallscale transient transfection and qiaprep spin miniprep kit qiagen, cat. The rneasy lipid tissue midi kit includes qiazol lysis reagent for lysing fatty do you have a protocol for the isolation of genomic dna andor proteins. Rna, proteins, dyes, and low molecular weight impurities are removed by a mediumsalt wash. Qiagen plasmid midi kit vs zymopure plasmid midiprep kit 25. For qiagen plasmid midi, maxi, mega, and giga kit protocols. Compared to the qiagen hi speed kit, i like both the invitrogen and qiagen plasmid plus more. Qiagen plasmid purification handbook qiagen plasmid midi, maxi, mega, and giga kits qiafilter plasmid midi, maxi, mega, and giga kits endofree plasmid maxi, mega, and giga kits for purification of ultrapure plasmid dna second edition august 2003.
Plasmid or cosmid dna purification using qiagen plasmid midi and maxi kits. Kits require use of a vacuum device for operation e. Qiaprep spin midimaxiprep kit protocol before staring make sure that rnase a has been added to p1 buffer. Qiagen plasmid maxi kit 10, 12162 from qiagen labsave. Should any product fail to perform satisfactorily due to any reason other. 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. Qiagen plasmid purification handbook harvard university. For the fastest, most convenient purification of up to 10 mg transfectiongrade plasmid dna. What is your preferred maxi prep kit for plasmid dna grown in. The procedure can be performed in 20 midi and maxi, 40 mega, or 50 minutes giga using a vacuum and centrifuge. The qiagen plasmid kits uses gravityflow qiagen anionexchange tips for efficient purification of plasmid dna. The qiagen plasmid plus kits enable ultrafast, largescale purification of up to 10 mg of highly pure plasmid dna. Mar, 2017 qiagen originals pure plasmids, genuine kits learn more.
Timerun, 100g yield, ideal for transfection, cloning, sequencing, lysate clearing without. Compactprep plasmid midi kit for fast purification of up to 200 g molecular biology grade plasmid dna. Qiagen plasmid plus purification handbook april 2012 en. For largescale genomic and viral dna purification from whole blood, plasma, serum, body fluids, lymphocytes. If using trishcl reagent, the qualities used should be recalculated. Qiagen plasmid plus kits provide a novel patentpending method for extremely. The use of a vacuum manifold allows purification of up to 24 samples in parallel, reducing handson time. Qiagen plasmid midi kit vs zymopure plasmid midiprep kit 25 preps this page will help you compare kit specs and protocols across manufacturers. The qiagen plasmid plus kits enable ultrafast, largescale purification of up to. For purifying plasmid dna from escherichia coli cells, the qiagen spin miniprep kit produces quite reliable results. 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 and drug. 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.