The PureLink™ HiPure Plasmid DNA Purification Kits use a patented anion-exchange resin to purify plasmid DNA to a level equivalent to two passes through CsCl gradients. The resin combines excellent capacity with a fast flow rate, high resolution, high yield, and efficient endotoxin removal. Plasmid preparations can typically be completed in less than 2 hours.

In the rare case that trace amounts of genomic DNA are still detectable in sensitive downstream applications such as e.g.,realtime RT-PCR, an in-solution digest using the RNase-Free DNAase set can be performed. Instructionsare presentedin Appendix C of the RNeasy MinElute Cleanup Handbook. In addition, tissue/cell lysis steps are typically carried out with lysis buffers containing guanidine isothiocyanate, a potent protein denaturant. It is very important to use a sufficient amount of lysis buffer during RNA isolation. We recommend using at least 10x volume of lysis buffer to tissue/cell pellet. CompactPrep Plasmid Kits enable fast, large-scale plasmid purification. The use of a vacuum manifold allows up to 24 samples to be purified in parallel on a single workbench (see figure "Microcentrifuge column format") for plasmid midi and plasmid maxi preps. Up to 200 µg plasmid DNA is purified in less than 60 minutes from 25 ml (high-copy plasmids) or 50 ml (low-copy plasmids) LB medium using the CompactPrep Plasmid Midi Kit. DNA is eluted in 100 µl and concentrations are typically 1–2 µg/µl. The CompactPrep Plasmid Maxi Kit enables purification of up to 750 µg plasmid DNA in less than 60 minutes from 100 ml (high-copy plasmid) or 250 ml (low-copy plasmid) LB medium. DNA is eluted in 200 µl and concentrations are typically 3–4 µg/µl. CompactPrep Plasmid Kits provide molecular biology grade plasmid DNA with low endotoxin levels (see figure "Low endotoxin levels"). Low yields of plasmid DNAcan be caused by a number of different factors. The most common causes for low yield are poor culturing conditions and plasmid propagation, excessive amounts of starting material resulting in insufficient bacterial celllysis and column overloading. When working with the anion-exchange based QIAGEN Plasmid Purification Kits, extra care is required during the isopropanol precipitation step, as the glassy DNA pellet may be difficult to see, and tends to be only loosely attached to the side of the tube. RNeasy Kits are the Gold standard for total RNA isolation. They provide fast purification of high-quality RNA from small to large amounts of cells, tissues, and yeast using silica-membrane RNeasy spin columns or 96-well plates. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor II or TissueLyser II or LT system. The RNeasy 96 Kit enables high-throughput purification of total RNA from up to 96 cultured-cell samples using silica-membrane RNeasy 96 plates. A dedicated RNeasy QIAcube Kit enables automated purification of 1–12 samples on the QIAcube Connect.On termination of the contract for any reason the customer shall immediately pay to VWR all of its outstanding unpaid invoices and interest. Confidentiality ensure complete disruption and homogenization of the starting material as instructed in the section 'Disruption and homogenization of starting materials' of the handbook

If VWR’s performance of the services is prevented or delayed by any act or omission of the customer, VWR shall without limiting its other rights or remedies, have the right to suspend performance of the services until the customer remedies the position and VWR shall not be liable for any losses or costs arising from such delay. Health, Safety and Liability If the cells in RNAprotect Tissue Reagent cannot be collected by centrifugation, please try one of the following suggestions: The level of endotoxin contamination in purified plasmid DNA depends on the purification method used (see table "Endotoxin levels in plasmid preparations"). Silica-slurry–purified DNA exhibits extremely high endotoxin levels. QIAGEN, QIAfilter, and HiSpeed Plasmid Kits and 2x CsCl ultracentrifugation yield very pure DNA with relatively low levels of endotoxin. EndoFree Plasmid Kits include an integrated endotoxin-removal step to yield plasmid DNA containing <0.1 EU/µg plasmid DNA. Customers who exceed their credit limits will be asked to pay in advance for additional products and/or services until the account is settled. Optimized buffers lyse samples, stabilize nucleic acids, and enhance selective DNA adsorption to the QIAamp membrane. Alcohol is added and lysates loaded onto the QIAamp spin column. Wash buffers are used to remove impurities and pure, ready-to-use DNA is then eluted in water or low-salt buffer. |Genomic DNA from 8 blood samples stored at 4°C for 1 week. DNA was purified using the QIAamp DNA Blood Mini Kit. When blood is stored at 4°C, the DNA is rapidly degraded due to apoptosis; the resulting apoptotic banding pattern can clearly be seen in these samples. M1: lambda-Hind III; M2:100 bp ladder.|Amplification of a 10 kb fragment of the human ALDH1 gene from genomic DNA isolated from blood. DNA was purified using conventional methods (Phenol) or the QIAamp DNA Blood Maxi Kit (QIAamp). PCR products were sequenced directly. M: 1 kb ladder.|Three paternity cases tested with DNA purified from blood samples with the QIAamp DNA Blood Mini Kit. 1: mother; 2: child; 3: alleged father; 4: child + alleged father. M: markers. Each lane had 3 µg of DNA loaded. Outcome: A: alleged father excluded; B & C: alleged father confirmed.

Achieve high-quality plasmid DNA with maxiprep extraction kits

Dilute the sample 10x by adding cold PBS. Pellet cells by centrifugation.Caution: Cells might lyse.

The concentration of RNA isolatedwith RNeasy Kits can be determined by measuring the absorbance at 260 nm (A260) in a spectrophotometer. Absorbance readings should be greater than 0.15 to ensure significance. An absorbance of 1 unit at 260 nm corresponds to 40 µg of RNA per ml (A260 = 1 = 40 µg/ml). This relationship is valid for measurements in water. Therefore, dilute RNA in waterto quantify it spectrophotometrically. Any additional or special terms included by VWR in its written acceptance shall form part of the contract. The terms and conditions of the contract apply equally to the supply of both products and services except where application to one or the other is specified. These terms will be governed by and construed in accordance with the laws of the State of Pennsylvania, without regard to any principles of conflicts of law. You agree that any action at law or in equity that arises out of or relates to these Terms and Conditions of Use will be filed exclusively in the state or federal courts located in Pennsylvania and you hereby consent and submit to the personal jurisdiction of such courts for the purposes of litigating any such action. The PureYield™ Plasmid Maxiprep System purifies up to 1mg of transfection-quality plasmid DNA from 250ml bacterial cultures in under an hour. The system is designed for use with a vacuum source and vacuum manifold, greatly reducing the time spent on purification compared to silica resin or other membrane-based methods. Toensure efficient gDNA removal when doing an on-column digest using the RNase-Free DNase Set in combination with RNeasy Minithe following factors are crucial:Any liability accepted by VWR under this contract is in lieu of any terms implied by law as to the quality or fitness for any particular purpose of the products and/or the standard of the services and all such implied terms are, to the fullest extent permitted by law, excluded from the contract between VWR and the customer. The customer shall indemnify VWR against any claims made against VWR by the customer’s employees, contractors or agents. Intellectual property rights Optionally, repeat the elution step, and incubate the spin column on the bench for 10 minutes with RNase-free water before centrifuging. VWR shall provide services to the customer in accordance with the specification agreed between them from time to time. Such services will be provided with all reasonable care and skill. The PureLink™ HiPure Plasmid Maxiprep Kit is designed to isolate transfection-grade plasmid DNA from E. coli. The Maxiprep Kit protocol will typically yield 500–850 µg plasmid DNA from 100–200 mL bacterial culture, at a purity that is comparable to that achieved by two passes through cesium chloride gradients.