Qiagen qiaprep spin miniprep kit, 250 preps, 1 to 100ml culture volume, 50l elution volume, molecular biology grade plasmid dna purification, silica technology, spin column format, manual processing, qiaprep spin miniprep kit. Four different cell lines cos7, hek293, hela, and huh7 were grown to 8090% confluence and transfected with 100 ng of each plasmid, in complex with 0. The genelute plasmid miniprep kit offers a simple, rapid, and costeffective method for isolating. Although many exceptions have been noted, the ribosomal rna fraction of the sample is more likely to represent either viable or. After dna extraction ive quantified the dna concentration and the result is a dna around 200ngul with. This tutorial explains how to carry out a miniprep using the qiaprep spin miniprep kit provided by qiagen. Kits can be stored for up to 12 months without showing any reduction in performance and qual ity. A majority of indoor residential microbes originate from humans, pets, and outdoor air and are not adapted to the built environment be. Plasmid dna encoding constitutively expressed gfp pegfpc2 was prepared using either monarch plasmid miniprep kit or qiagen qiaprep spin miniprep kit. How to do an endofree maxi prep thermo fisher scientific. Contact qiagen technical services or your local distributor for protocols for these applications. Plasmid dna purification using qiaprep spin miniprep kit. This protocol is designed for the purification of up to 20 ug highcopy plasmid dna from 15 ml overnight e.
The final protocol for large scale mirapreps was tested with genejet, qiagen, and sigma genelute spin column miniprep kits, using the resuspension, lysis, neutralization and wash buffers provided a stepby step protocol is provided in s1 file. This represents a time saving of 50 % compared with qiaprep spin miniprep kit. Qiagen qiaprep spin miniprep column, 50l elution volume, 1 to 100ml culture volume, 1 to 24 samplesrun. For small plasmids, lets say below 50 kb, you can try one of the several commercial kits. Both the qiaprep spin miniprep kits and qiagen plasmid plus kits work with the manifold, which can handle up to 24 samples at the. Plasmid isolation from yeast using qiagen qiaprep kit. We have started using the zymoprep yeast plasmid miniprep kit ii with much success. For purification of up to 20 micro g molecular biology grade plasmid dna. The unique spin column design also provides zero buffer retention and a low elution volume. Qiaprep spin miniprep kit 50 laboratory consumables. Kit contents qiaprep spin miniprep kit 50 250 catalog no. Consequently, a large portion of the microbes identified by dnabased methods are either dead or metabolically inactive. 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. I used for the first time the qiagen plasmid miniprep kit to extract dna from my plasmids.
Qiagen qiaprep spin miniprep kit, 50 preps, 1 to 100ml culture volume, 50l. Qiaprep spin miniprep kit highyield protocol english pdf. Pellet cells in 15 ml falcon tube, resuspend in water, and transfer to 1. Qiaprep spin miniprep kit protocol for yeast using a microcentrifuge procedure 1. For plasmids reaching sizes of 50 kb you may look for cosmid dna isolation when looking for a suitable. Plasmid dna purification using the qiaprep spin miniprep. The standard protocol uses 50 l buffer eb for elution, since this combines high yield. Resuspend pelleted bacterial cells in 250ul buffer p1 and transfer to a microcentrifuge tube. Dna, rna, viral nucleic acids and proteins, plus dna and rna cleanup. 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, kit, qiaprep spin miniprep. The qiaprep spin miniprep kit is designed for isolation of up to 20. Qiaprep spin miniprep kit 50, 27104 from qiagen labsave. Qiaprep miniprep handbook washington state university. The qiaprep spin miniprep kit is designed for quick and convenient processing of 124 samples simultaneously in less than 30 minutes.
The qiacube connect is preinstalled with protocols for purification of plasmid dna, genomic. Timerun, 20g yield, silica technology, manual processing, ideal for ligation, cloning, transformation etc. Data file 28907592 aa plasmid dna purification illustra. We have had good success with this kit the instructions are very easy to follow and all the necessary reagents are supplied buffer p1 must be. The binding capacity of the columns within the miniprep kit is 20 ug of plasmid dna. Qiaprep spin miniprep kit for purification of high quality plasmid dna. All steps are performed without the use of phenol, chloroform, cscl, ethidium bromide, and without alcohol precipitation. Qiaprep spin miniprep kit 250 from qiagen sample to.
Qiaprep spin miniprep kit 50 from qiagen selectscience. The most difficult buffer for miniprep is the neutralization buffer columns. Centrifuge for 1 min to remove residual wash buffer. The kit comes with all components necessary for cell lysis, silicagel spin columns, high and low salt buffers and wash buffers to perform plasmid dna purification. Plasmid dna purification using the qiaprep spin miniprep kit. Purifying plasmid dna using the qiagen miniprep kit protocol. Even higher yields with the highyield supplementary protocol. Qiaprep spin miniprep kit 50 from qiagen sample to insight. Timerun, 20g yield, ideal for fluorescent and radioactive sequencing. The classic qiagen bindwashwashelute protocol for the qiaprep spin miniprep kit involves several centrifugation steps. Genejet plasmid miniprep kit utilizes an exclusive silicabased membrane technology in the form of a convenient spin column. We have experience using the qiaprep spin miniprep kit, which can be used to process 120 samples in about 3045 minutes. Qiaprep spin miniprep kit from qiagen biocompare product. Qiaprep spin miniprep kit protocol using 5ml collection tubes the qiaprep spin miniprep procedure can be performed using 5ml centrifuge tubes.
Thermo scientific genejet plasmid miniprep kit for 50. Comparison of the final miraprep protocol to commercial silica spin column dna preparations. 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. A comparison of methods used to unveil the genetic and. Buffer composition for miniprep kit molecular biology. The kits can be purchased in quantities of 50 250 columns. The kit utilizes an exclusive silicabased membrane technology in the form of a convenient spin column. Genelute spin column miniprep kits, using the resuspension, lysis, neutralization and wash buffers provided a stepby step protocol is provided in s1 file. Grow yeast strain containing desired plasmid overnight in 10 ml of rich media yepd. With a simple protocol, fast handling and reproducible results, what else could you need.