Highlights efficienthigh yields of up to 20 g of high quality plasmid dna fastpr. Qiagen qiaprep spin miniprep kit, 50 preps, 1 to 100ml culture volume, 50l elution volume, molecular biology grade plasmid dna purification, silica technology, spin column format, manual processing. Remove the spin filter from the 2 ml tube, discard the filtrate at. If you have never done this protocol before, read the the background information in the handbook like the important notes section.
Background information 29 growth of bacterial cultures 29 preparation of cell lysates 32. Buffer composition for miniprep kit molecular biology. Compare qiaprep spin miniprep kit from qiagen sample to insight on catalog number. Thermo scientific genejet plasmid miniprep kit for 50. Qiaprep spin 22 qiaprep miniprep handbook 122006 protocol. The following has been reproduced from the handbook and annotated based on experience with the kit. This tutorial explains how to carry out a miniprep using the qiaprep spin miniprep kit provided by qiagen. Quantum prep plasmid miniprep kit instruction manual. The most difficult buffer for miniprep is the neutralization buffer columns.
The monarch plasmid miniprep kit is a rapid and reliable method for the purification of high quality plasmid dna. Qiaprep spin miniprep kit was used to purify dna from 1. Qiaprep miniprep handbook 052004 3 contents kit contents 4 storage 7 quality control 7 product use limitations 7 product warranty and satisfaction guarantee 8 technical assistance 8 safety information 9 introduction 10 principle qiavac vacuum manifolds 16 guidelines for qiavac manifolds 18 important notes 19 protocols qiaprep spin miniprep kit. 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. Purifying plasmid dna from bacterial colonies using the qiagen miniprep kit. This is the protocol for qiagens qiaprep spin miniprep kit catalog numbers 27104 or 27106 with their newer version 2. Data file 28907592 aa plasmid dna purification illustra. Research open access identification of a histidine metal. Timerun, 20g yield, ideal for fluorescent and radioactive sequencing. Plasmid dna purification using the qiaprep spin miniprep kit and a.
Qiaprep miniprep handbook 122006 5 qiaprep 8 turbo miniprep kit 10 50. The qiaprep spin miniprep kit is designed for quick and convenient processing of 124 samples simultaneously in less than 30 minutes. The classic qiagen bindwashwashelute protocol for the qiaprep spin miniprep kit involves several centrifugation steps. Use of the recommended lb composition with 10 gliter nacl, also see appendix a, p. Qiaprep spin miniprep kit using a microcentrifuge this protocol is designed for purification of up to 20 g of highcopy plasmid dna from 15 ml overnight cultures of e. Objectives after completion, the student should be able to. Qiaprep miniprep handbook university of pittsburgh. The qiaprep spin miniprep kit can be fully automated on the qiacube connect.
The qiaprep spin miniprep kit is designed for isolation of up to 20. While guaranteeing excellent flowthrough of all the wash buffers and. Thermo scientific genejet plasmid miniprep kit for 50 preps life sciences. Resuspend pelleted bacterial cells in 250 ul buffer p1 and transfer to a microcentrifuge tube. Grow yeast strain containing desired plasmid overnight in 10 ml of rich media yepd. Features and benefits from harvested bacterial culture to pure plasmid dna in 30 minutes or less up to 350.
Grow yeast strain containing desired plasmid overnight in 10 ml of rich. Ultraclean6 minute mini plasmid prep kit is used by thousands of researchers all over the world. Enzymatic assay of arge arge activity was measured by the method of javidmajd and blanchard javidmajd and blanchard 2000. Genelute plasmid miniprep kit offers a simple, rapid, and costeffective method for isolating up to 20 g plasmid dna from e. This method employs standard cell resuspension, alkaline lysis, and neutralization steps, with the additional benefit of color indicators at certain steps to easily monitor completion. Qiaprep spin qiaprep miniprep handbook 052004 23 protocol.
Highsalt buffer is added to neutralize the lysate and to create appropriate conditions for dna binding to the silica membrane. Pdf plasmid purification is a basic tool of molecular biologists. 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. The kit recovers up to 20 g of high copy plasmid dna per isolation procedure. Jul 20, 2015 this tutorial explains how to carry out a miniprep using the qiaprep spin miniprep kit provided by qiagen. Kit contents qiaprep spin miniprep kit 50 250 catalog no. Part of the zymopure plasmid kits collection, the zymopure plasmid miniprep kit features a spin columnbased method for the purification of up to 100 g of ultrapure endotoxinfree plasmid dna in less than 15 minutes. Protocol is designed for the purification of up to 20. Qiagen research kits compatible with the qiavac 24 plus application kit accessories plasmid minipreps qiaprep spin miniprep kits dna cleanup qiaquick pcr purification kits qiaquick gel extraction kits qiaquick nucleotide removal kits minelute pcr purification kits. Plasmid dna purification using the qiaprep spin miniprep.
Bacterial cells are harvested via centrifugation, subjected to a modified alkalinesds lysis. The qiaprep miniprep system is intended for isolation of plasmids from bacteria cells. Q i a p r e p s p i n qiaprep miniprep handbook 052012 19 protocol. After centrifugation, the clear supernatant is loaded onto a nucleospin plasmid miniprep spin column. Qiaprep spin columns can be used in a microcentrifuge or on any vacuum manifold with luer connectors e. Qiaprep spin midimaxiprep kit protocol before staring make sure that rnase a has been added to p1 buffer. Qiagen qiaprep spin miniprep kit 27104 or 27106 with a. Qiaprep spin miniprep kit protocol for yeast using a microcentrifuge. The kit uses silicagelmembrane technology to bind dna, from which the dna can be eluted with an elution buffer or water. The kit utilizes an exclusive silicabased membrane technology in the form of a convenient spin column. Qiaprep spin miniprep kit protocol using 5ml collection tubes the qiaprep spin miniprep procedure can be performed using 5ml centrifuge tubes e. Genelute hp plasmid dna midiprep kit 25 purifications gen. Qiaprep spin miniprep kit protocol using a microcentrifuge. Plasmid dna purification using the qiaprep spin miniprep kit and a microcentrifuge this protocol is designed for purification of up to 20.
Plasmid dna purification using the qiaprep spin miniprep kit and a microcentrifuge the idea a miniprep is a way of extracting plasmid dna from cells. The qiaprep spin miniprep kit can be fully automated on the qiacube. Qiaprep spin miniprep kit from qiagen biocompare product. Qiaprep spin qiaprep spin miniprep kit protocol using a vacuum manifold this protocol is designed for purification of up to 20 g highcopy plasmid dna from 15 ml overnight cultures of e. Qiaprep spin miniprep kit highyield protocol english pdf. Pureyieldplasmid miniprep system instructions for use of products a1220, a1221, a1222 and a1223 protocol quick alternative protocol for larger culture volumes 1. Purifying plasmid dna using the qiagen miniprep kit protocol. Plasmid dna purification using qiaprep spin miniprep kit. Pellet cells in 15 ml falcon tube, resuspend in water, and transfer to 1.
Qiaprep spin miniprep kit 50 from qiagen selectscience. Thermo scientific genejet plasmid miniprep kit utilizes an exclusive silicabased membrane technology in the form of a convenient spin column. Qiaprep miniprep handbook 112005 7 storage qiaprep miniprep kits should be stored dry at room temperature 1525c. Genelute hp plasmid dna midiprep kit 25 purifications. Qiaprep spin miniprep kit protocol for yeast using a microcentrifuge procedure 1.
Typically the user of qiaprep must cut off tube caps to use the qiaprep kit 6. Plasmid dna purification using the qiaprep spin miniprep kit and a microcentrifuge 19 and 5 ml collection tubes 20 and a vacuum manifold 21 plasmid dna purification using the qiaprep 96 turbo miniprep kit 23 troubleshooting guide 26 appendix a. Spin 2 ml of overnight culture for 30 seconds spin 2 ml of culture. This protocol is designed for purification of up to 20. Apr 28, 2014 the classic qiagen bindwashwashelute protocol for the qiaprep spin miniprep kit involves several centrifugation steps.
Plasmid dna miniprep student guide goal the objective of this lab is to perform extraction of plasmid dna and analyze the results. Low throughput the qiaprep spin miniprep kit is designed for quick and convenient processing of 124 samples simultaneously in less than 30 minutes. Qiaprep miniprep handbook washington state university. Harvested bacteria are resuspended and processed by sdsalkaline lysis. A protocol that uses a miniprep kit and provides maxiprep yields. The unique spin column design also provides zero buffer retention and a low elution volume. Genelute hp plasmid midiprep kit has been used to isolate and purify plasmids.
Plasmid dna purification using the qiaprep spin miniprep kit and a microcentrifuge this protocol is designed for purification of up to 20 g of highcopy plasmid dna from. The innovative qiacube uses advanced technology to process qiagen spin columns, enabling. Qiaprep spin miniprep kit highyield protocol english pdf print bookmark share for purification of up to 30. Qiagen is the leading provider of innovative sample and assay technologies, enabling plasmid dna purification using the qiaprep spin miniprep kit. The standard protocol on pages 1819 should be followed with the following.
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