Mirapreps and minipreps were conducted using either genejet from thermo fisher waltham, ma, usa, qiagen hilden. Qiaprep spin miniprep kit highyield protocol english pdf for purification of up to 30. The following procedure is performed at room temperature. The range of protocols available is continually expanding, and additional qiagen protocols can. Dna of up to 10 kb is purified using a simple and fast bindwashelute procedure and an elution volume of 3050. Find out how to transfer protocols to your qiacube. Qiaprep miniprep kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Pure plasmid dna can improve your automated sequencing results by producing.
Qiaprep spin miniprep kit using a microcentrifuge 23 using 5 ml collection tubes 24 using a vacuum manifold 25 qiaprep 8 miniprep kit 28 qiaprep 8 turbo miniprep kit 31 qiaprep 96 turbo miniprep kit 33 cell cultivation in a 96well block 36 special applications 37 troubleshooting guide 38 appendix a. For more information, please refer to the qiaprep miniprep handbook, december. Gencatchtm plus plasmid dna miniprep kit provides a simple, fast and. Protocols are fast and user friendly, saving you valuable time. Tri reagent is provided with catalog numbers r2071 and r2073 only. For higher yields and purity use the alternative protocol below to harvest and process up to 3ml of bacterial culture. The endofree maxiprep kit from qiagen or the genejet maxiprep kit.
Genelute plasmid dna miniprep kit 70 purifications gen. The qiaprep spin miniprep kit is designed for quick and convenient processing of 124 samples simultaneously in less than 30 minutes. Quick start procedure this quick start procedure is for experienced users only. Quickstart protocol february 2015 qiaprep spin miniprep kit the qiaprep spin miniprep kit cat. Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. Add the provided rnase a solution to buffer mx1, mix and store at 4 c. Each amplification curve represents an average of three independent dnafree rna the quickrna miniprep plus kit is universal and accommodates any sample input including cells, tissue, blood etc. Qiaprep miniprep handbook 052012 5 storage qiaprep miniprep kits should be stored dry at room temperature 1525c.
Total rna was isolated from human epithelial cells sans dnase treatment. A link to the relevant protocol sheet is provided in the detailed information about the protocol. This is the protocol for qiagen s qiaprep spin miniprep kit catalog numbers 27104 or 27106 with their newer version 2. They are widely used in many laboratories, but core principle is based on. Whats the difference between the qiagen spin miniprep kit, and the. Plasmid dna miniprep kits thermo fisher scientific us. Qiaprep miniprep handbook 052004 7 storage qiaprep miniprep kits should be stored dry at room temperature 1525c. Thermo scientific genejet plasmid miniprep kit has been tested in the isolation of puc19 dna from e. The protocol presented in this technical bulletin is for the isolation of plasmid dna from e.
The qiaprep spin miniprep kit is designed for isolation of up to 20. The qiacube is preinstalled with protocols for purification of plasmid dna, genomic dna, rna, viral nucleic acids, and proteins, plus dna and rna cleanup. Ensure that buffers have been prepared according to the instructions on page 3. The monarch total rna miniprep kit is a comprehensive solution for sample preservation, cell lysis, gdna removal, and purification of total rna from a wide variety of biological samples, including cultured cells, blood, and mammalian tissues. Plasmid dna can be purified from 110ml of overnight cultures of e.
For purification of total rna including small rnas 50 g, the quickrna plant miniprep kit features a specially formulated rna lysis buffer. Objectives after completion, the student should be able to. General description the genelute plasmid miniprep kit offers a simple, rapid, and costeffective method for isolating plasmid dna from recombinant e. Qiaprep miniprep handbook 122006 7 storage qiaprep miniprep kits should be stored dry at room temperature 1525c. The quicklyse miniprep kit provides a rapid spincolumnbased method for isolating sequencinggrade plasmid dna. The protocol describes the preliminary harvesting of bacteria and incubation with lysozyme to lyse their cell walls. Jul 20, 2015 this tutorial explains how to carry out a miniprep using the qiaprep spin miniprep kit provided by qiagen.
This new plasmid dna isolation protocol will significantly reduce time. You can search for protocols and protocol sheets by selecting an application, kit, or starting material. Qiaquick spin kits are tested by isolation of dna fragments of various sizes from either aqueous solution or agarose gel. Qiagen robotic systems are not available in all countries, please inquire. Qiaprep spin miniprep kit for purification of high quality plasmid dna.
A the zymobiomics dna miniprep kit provides unbiased representation of the organisms extracted from the zymobiomics microbial community standard. As part of the stringent qiagen quality assurance program, the performance of qiaquick spin kits is monitored routinely and on a lottolot basis. Whole blood fresh or stored, buffy coat, buccal cells, cells from culture, saliva, and other biological liquid samples can be processed with these kits. Add 40 ml 50 preps or 180 ml 250 preps 98100% ethanol. The kit recovers up to 20 g of high copy plasmid dna per isolation procedure. Plasmid dna miniprep student guide goal the objective of this lab is to perform extraction of plasmid dna and analyze the results. The protocol describes the preliminary harvesting of bacteria and incubation with lysozyme to lyse their cell walls before dna purification. Here, we videorecorded the general procedures of miniprep through the qiagen s qiaprep 8 miniprep kit, aiming to introducing this highly efficient technique to the general beginners for molecular biology techniques.
Qiaprep miniprep handbook washington state university. Additionally, bacteria, yeast, and plant samples can be processed with. Plasmid dna purification using the qiaprep spin miniprep kit and a. The genelute mammalian genomic dna miniprep kit provides a simple and convenient way to isolate pure genomic dna from a variety of cultured cells, tissues including rodent tails, and fresh whole blood or white blood cells. Lyophilized qiagen protease can be stored at room temperature 1525c for up to. The quality of isolated dna has been evaluated in a set of tests. A protocol that uses a miniprep kit and provides maxiprep yields. Qiaprep miniprep handbook 122006 3 contents kit contents 4 storage 7 quality control 7 product use limitations 7 product warranty and satisfaction guarantee 7 technical assistance 8 safety information 9 introduction 10 principle 11 using lyseblue reagent 14 important notes 15 guidelines for qiavac manifolds 18 protocols. Qiaprep spin miniprep kit protocol using 5ml collection tubes the qiaprep spin miniprep procedure can be performed using 5ml centrifuge tubes e. Dna was extracted from zymobiomics microbial community standard using four different dna extraction methods zymobiomics dna miniprep kit, human microbiome project protocol, supplier q1, and. Most widely accepted protocol for transfection rapid purification with integrated lysate clarification filter rapid, highthroughput transfectiongrade purification. These are available online in convenient and compact pdf format at. The yield of plasmid will vary depending on a number of factors, including the volume of bacterial culture.
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. The quickrna plant miniprep kit provides for rapid isolation of rna from various plant samples e. Highlights efficienthigh yields of up to 20 g of high quality plasmid dna fastpr. The procedure combines a unique buffer system with zymospin column technology to yield high quality total rna including small rnas 17200 nt in about 10 minutes. First time users are strongly recommended to read through the detailed instruction protocol in section 4. For technical assistance, please call tollfree 0080022446000, or find regional phone numbers. Miniprep kit is a plasmid dna purification system designed to yield pure plasmid dna figures 1 and 2 for automated sequencing in just 25 minutes. The standard protocol on pages 2223 should be followed with the following modifications. Find protocols, product documentation, software, faqs, videos, webinars, and more.
The qiaprep spin miniprep kit can be fully automated on the qiacube connect. The quickdna kits are ideal dna isolation kits for easy, rapid isolation of total dna e. The quickrna miniprep kit is one of the most innovative rna isolation kits available, designed for the easy, reliable, and rapid isolation of dnafree rna from a wide range of cell up to 10 7 and tissue samples up to 50 mg. Purifying plasmid dna using the qiagen miniprep kit protocol. The genelute plasmid miniprep kit offers a simple, rapid, and costeffective method for isolating plasmid dna from recombinant e. 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. Qiaquick pcr purification kit protocol using a microcentrifuge this protocol is designed to purify single or doublestranded dna fragments from pcr and other enzymatic reactions see page 8. The quickrna miniprep plus kit is one of the most innovative rna isolation kits available, designed for the easy, reliable, and rapid isolation of dnafree rna from cells, all tissue types, whole blood, and biological fluids. C plasmid preparations with the genelute kit, comparing either the standard miniprep procedure or the miraprep using 1x volume of ethanol. These are available online in convenient and compact pdf. Longer reads lower background higher accuracy stronger signal.
Qiagen plasmid purification handbook harvard university. Product information thermo scientific genejet plasmid. Total rna was isolated from 106 human epithelial cells with incolumn dnase treatment for both kits. For purification of ultrapure, transfectiongrade plasmid dna. Spin miniprep kit for purification of highquality plasmid dna. Wizard plus sv minipreps dna purification system protocol. Quickstartprotocol qiaprep spin miniprep kit october 2010. They can be stored for at least 2 years without showing any reduction in performance, capacity, or quality of separation. The range of protocols available is continually expanding, and additional qiagen protocols can be. Qiagen plasmid purification handbook home facebook. The standard protocol on pages 1819 should be followed with the following modifications. The collaborative platform and preprint server for science methods and protocols, computational workflows, clinical trials, operational procedures, safety check qiagen qiaprep spin miniprep kit 27104 or 27106 with a microcentrifuge. The quickrna miniprep kit is one of the most innovative rna isolation kits available, designed for the easy, reliable, and rapid isolation of dnafree rna from a wide range of cell up to 107 and tissue samples up to 50 mg.
Dna purification by centrifugation prepare lysate 1. The qiaprep spin miniprep kit can be fully automated on the qiacube. Should i use the qiagen spin miniprep kit, or the qiagen plasmid mini kit for my plasmid. More information about biocoder can be found on my home page. In accordance with qiagen s 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. Qiaprep spin miniprep kit highyield protocol english pdf. Qiagen is the leading provider of innovative sample.
Perform steps 18 as indicated in the general protocol, see page 4. Quick start procedure this quick start procedure is for. The procedure combines a unique buffer system with zymospin column technology to yield hig. Qiaamp dna mini kit and qiaamp dna blood mini kit handbook. Our product finder helps you select the best kit for your needs use productfinder.
Under these conditions, if no expiration date is mentioned on the kit label, qiaprep miniprep kits can be stored for up to 12 months without showing any reduction in performance and quality. The provided dnarna shield stabilizes samples, allowing them to be stored without the need f. For uptodate licensing information and productspecific disclaimers, see the respective qiagen kit. Plasmid dna purification using the qiaprep spin miniprep kit. By combining silicabinding technology and the convenience of a spin column format, up to 15. Genelute mammalian genomic dna miniprep kit protocol. 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. The ultrafast purification protocol, based on the remarkable selectivity of patented qiagen resin, allows the. Detailed protocols for dna and plasmid extraction procedures. The quickrna miniprep kit yields high quality total rna. For a more refined search, select more than one parameter.
Qiaprep spin miniprep kit protocol using 5 ml collection tubes the qiaprep spin miniprep procedure can be performed using 5 ml centrifuge tubes e. Hispeed plasmid purification handbook harvard university. Qiagen plasmid purification handbook 082003 9 product use limitations qiagen, qiafilter, and endofree plasmid kits are developed, designed and sold for research purposes only. List the function of each solution used in a plasmid dna miniprep. What you see here is the autogenerated text ouput of the protocol that was coded up in biocoder see source code. Qiaprep miniprep handbook 112005 7 storage qiaprep miniprep kits should be stored dry at room temperature 1525c. For qiagen plasmid midi, maxi, mega, and giga kit protocols. Equal volumes of eluted dna were then analyzed in duplicate in a. Please consult the material safety data sheet for information regarding hazards and safe handling practices. Following is the miniprepqiagen kit protocol in biocoder, a highlevel programming language for expressing biology protocols.
Kits can be stored for up to 12 months without showing any reduction in performance and qual ity. Under these conditions, if no expiration date is mentioned on the kit label, qiaprep miniprep kits can be stored for up to 12 months without showing any reduction in. Thermo scientific genejet plasmid miniprep kit utilizes an exclusive silicabased membrane technology in the form of a convenient spin column. The monarch plasmid miniprep kit is a rapid and reliable method for the purification of up to 20. For cleanup of other enzymatic reactions, follow the protocol as described for pcr samples or use the minelute reaction cleanup kit. 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. This protocol is designed for purification of total dna from grampositive bacteria. Kits can be stored for up to 12 months without showing any reduction in performance and quality.
The qiaquick pcr purification kit provides spin columns, buffers, and collection tubes for silicamembranebased purification of pcr products 100 bp. Innovation by qiagen creates sample and assay technologies that enable insights making improvements in life possible. Qiagen qiaprep spin miniprep kit 27104 or 27106 with a. Pdf plasmid purification is a basic tool of molecular biologists.