Plasmid mini kit qiagen protocol pdf

Plasmid dna mini kit i protocol spin protocol all centrifugation should be performed at room temperature unless otherwise noted. These miniprep columns contain a unique silica membrane that binds up to 50 g of plasmid dna. Qiagen, qiafilter, hispeed, and endofree plasmid kits, provide optimal. 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 5 kit contents qiagen plasmid kits qiagen plasmid kit midi 25 midi 100 maxi 10 maxi 25 maxi 100 catalog no. Add 10 ml of buffer p3, mix gently, incubate on ice for 1520 min. For qiagen plasmid midi, maxi, mega, and giga kit protocols. Qiaprep miniprep handbook for purification of molecular biologygrade dna plasmids large plasmids 10 kb. For purification of lowcopy plasmids and cosmids, large plasmids 10 kb, and dna prepared using other. Qiaprep miniprep handbook 112005 3 contents kit contents 4 storage 7 quality control 7. Qiaamp dna mini kit and qiaamp dna blood mini kit handbook. Recovering plasmid dna from bacterial culture addgene.

Forceps are to be sterilized in fine science tools heat block at. These are available online in convenient and compact pdf format. We dramatically improved a plasmid isolation protocol based on the popular. Qiagen plasmid purification handbook sierra bermeja. Important notes the yield of plasmid dna using the miniprep kit depends on. This plasmid kit will purify plasmid dnacosmid dna from 15 ml of cultured bacterial cells using an efficient plasmid miniprep system. Qiaprep miniprep handbook washington state university. The range of protocols available is continually expanding, and additional qiagen protocols can. Qiagen qiaprep spin miniprep kit 27104 or 27106 with a. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. The protocol for hispeed midi and maxi kits is optimized for use with cultures grown. The range of protocols available is continually expanding, and additional qiagen protocols can be. Qiagen hispeed plasmid midi kit, 25 preps, 50l to 150ml culture volume, transfectiongrade plasmid or cosmid dna purification, anionexchange technology, manual processing, english pdf.

Qiagen buffer also works for epoch life sciences spin columnswhich are sold in bulk at a much lower price. Qiagen plasmid purification handbook kirschner lab. The rneasy mini kit, rneasy protect mini kit including rnalater rna. For purifying plasmid dna from escherichia coli cells, the qiagen spin miniprep kit produces quite reliable results. This kit contains all the re agents necessary for the purification of plasmid dna by alkaline lysis and alcohol precipitation. Qiagen robotic systems are not available in all countries, please inquire. 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. No claim or representation is intended for their use to provide information for the diagnosis, prevention, or treatment of a disease.

The kit recovers up to 20 g of high copy plasmid dna per isolation procedure. For uptodate licensing information and product specific disclaimers, see the respective qiagen kit handbook or user manual. For uptodate licensing information and productspecific disclaimers, see the respective qiagen kit handbook or user manual. This protocol is designed to isolate plasmid dna from e. Protocols plasmid or cosmid dna purification using qiagen plasmid midi and maxi kits 16. Highlights efficienthigh yields of up to 20 g of high quality plasmid dna fastpr. These are available online in convenient and compact pdf format at. Qiagen plasmid kit midi 25 midi 100 maxi 10 maxi 25 maxi 100 catalog no.

Qiaprep spin midimaxiprep kit protocol before staring make sure that rnase a has been added to p1 buffer. This kit can be used to purify plasmids from the colonies generated with the bacterial transformation kit cat. Plasmid dna purification using the qiaprep spin miniprep kit and a. Qiagen provides a variety of qiaamp genomic dna kits for fast, efficient and. If you want to perform plasmid purification without using a kit, you can find a protocol for kit free plasmid mini prep at the bottom of this page. We design our plasmid dna extraction products to isolate plasmid dna at the purity and scale you need. Mix gently by inverting the tube 46 times and incubate at room temperature for exactly 5 min. Product information thermo scientific genejet plasmid. Spin miniprep kit for purification of highquality plasmid dna. The purchaser must determine the suitability of the product for its particular use. Qiaprep miniprep handbook 052012 3 contents kit contents 4 storage 5 intended use 5 safety information 5 quality control 6 introduction 7 principle 8 using lyseblue reagent 11 important notes 12 guidelines for qiavac manifolds 15 protocols plasmid dna purification using the qiaprep spin miniprep kit and a microcentrifuge 19 and 5 ml collection. Please consult the material safety data sheet for information regarding hazards and safe handling practices. The protocol below is meant to describe the general procedure for purifying plasmid dna from bacterial cultures.

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. For purification of lowcopy plasmids and cosmids, large plasmids 10. For qiagen buffer compositions, please see the qiagen buffers page. For cosmid and lowcopynumber plasmid purification see recommendations on page 12. Hispeed plasmid midi kit 25 from qiagen sample to insight. The nucleospin plasmid miniprep kit is designed for the rapid 25 minutes, highyield up to 45 g, smallscale preparation of highpurity plasmid dna. Choose from our wide range of highperforming, costeffective plasmid purification technologies designed to help you overcome common plasmid prep issues, such as low recovery and the presence of impurities. Should any product fail to perform satisfactorily due to any reason other. The protocol describes the preliminary harvesting of bacteria and incubation with lysozyme to lyse their cell walls before dna purification. Qiaprep spin miniprep kit for purification of high quality plasmid dna.

The qiacube is preinstalled with protocols for purification of plasmid dna, genomic dna, rna, viral nucleic acids, and proteins, plus dna and rna cleanup. Our technical service departments are staffed by experienced scientists with extensive practical and theoretical expertise in molecular biology and the use of qiagen prod ucts. The qiacube is preinstalled with protocols for purification of plasmid dna, genomic. 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. Isolate a suitable piece of tissue and place in a uvcrosslinked 1. Qiagen plasmid mini handbook 0399 9 protocol plasmid mini purification protocol this protocol is for mini up to 20 g preparations of highcopy plasmid dna from cultures of e. Product use limitations the mirneasy mini kit is intended for research applications. Jul 20, 2015 this tutorial explains how to carry out a miniprep using the qiaprep spin miniprep kit provided by qiagen. Choose from our wide range of highperforming, costeffective plasmid purification technologies designed to help you overcome common plasmid prep issues. Follow the qiagen protocol if there is confusion or a discrepancy. This is the protocol for qiagens qiaprep spin miniprep kit catalog numbers 27104 or 27106 with their newer version 2. We dramatically improved a plasmidisolation protocol based on the popular.

Qiagen plasmid purification handbook harvard university. For purification of bacs, pacs, p1s, and purification of double. The quality of the presto mini plasmid kit is tested on a lottolot basis according to geneaids isocertified quality management system. Important notes 15 guidelines for qiavac manifolds 18 protocols plasmid dna purification using the qiaprep spin miniprep kit and a microcentrifuge 22 and 5 ml collection tubes 23 and a vacuum manifold 24. If you will be using a kit, follow the kits instructions. Purifying plasmid dna using the qiagen miniprep kit protocol. Plasmid dnas purified from this kit can be used directly for downstream applications such as transfection, cloning, sequencing, as well as in vivo applications. 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. Mirapreps and minipreps were conducted using either genejet from thermo fisher waltham, ma, usa, qiagen hilden. Add 10 ml of buffer p3, mix gently, and incubate on ice for 1520 min. Qiagen plasmid purification handbook qiagen plasmid midi, maxi, mega, and giga kits. Qiaprep spin miniprep kit highyield protocol english pdf.

Plasmid purification mini kit protocol using a microcentrifuge please read important notes on pages 4 before starting. These are available online in convenient and compact pdf. If you will be using a kit, follow the kit s instructions. The qiaprep spin miniprep kit is designed for isolation of up to 20. If you wish to process larger volumes of bacterial culture up to 3ml use the alternative protocol provided below. For the qiagen plasmid mini kit protocol microcentrifuge 1. The qiacube connect is preinstalled with protocols for purification of plasmid dna, genomic dna, rna, viral nucleic acids and proteins, plus dna and rna cleanup.