Conventional breeding of switchgrass biomass is difficult because it displays selfincompatible hindrance. Homogenates were subjected to a single freezethaw cycle to aid cell lysis and then cleared by centrifugation at 10,000 x g for 5 min at 4 c. Up to 400 mg agarose can be processed per spin column. Samples were homogenised using a tissuelyser ii qiagen with 5 mm stainless steel beads for 3. Alternatively, a grinding jar set can be used to process large samples. Dneasy blood and tissue kit 50 from qiagen sample to. Therefore, direct genetic modifications of switchgrass have been considered the more effective approach to tailor switchgrass with traits. Aug 24, 2018 a detailed investigation of the porcine skin and nose microbiome using universal and staphylococcus.
The tissuelyser enables fast and uniform disruption of animal and human tissues, plant tissues, bacteria, and yeast in various sample volumes in several formats. The log file is updated as new runs are carried out, with the oldest data being deleted. Qiagen tissueruptor ii sample disruption tools see details. All beadbeating steps were performed in the qiagen tissuelyser ii at a frequency of 30 hz for 2 min and centrifugation steps were carried out in an eppendorf 5415d centrifuge.
Lt is comparable to that achieved with the wellestablished tissuelyser ii figures 6 and 810. Qiacube ht laboratory equipment pdf manual download. For disruption of up to 48 or 192 biological samples. Qiaquick gel extraction kit protocol using a microcentrifuge.
If the tissuelyser ii is not in use, switch it off at the power switch. Microbiome dysbiosis is associated with disease duration. Rna preservation agents and nucleic acid extraction method. For disruption of up to 48 or 192 biological samples 48 pages. Qiagen tissuelyser ii sample disruption 11843 youtube. Forceps are to be sterilized in fine science tools heat block at. Servicing of the tissuelyser ii should only be performed by qiagen or an authorized agent. Evaluating variation in human gut microbiota profiles due to.
A molecular clone of chronic bee paralysis virus cbpv. Marked strainspecific differences in the su5416 rat model. Rna extraction was performed using the qiagen rneasy fibrous tissue mini kit run on the qiacube qiagen. It ramps up to 30 shakes per second and the timer works as it should. Add the appropriate volume of buffer rlt see table 8, and homogenize by passing the lysate at least 5 times through a blunt 20gauge needle fitted to an rnasefree syringe. Isolate a suitable piece of tissue and place in a uvcrosslinked 1. Gel extraction qiagen qiaquick gel extraction kit materials. Protein lysate was quantified using the bca bicinchoninic acid assay method. Do not continue to use the tissuelyser ii or tissuelyser accessory if it is damaged. For highly specific and sensitive multiplex pcr without optimization requirements. Jul 07, 2017 qiagen tissuelyser ii sample disruption 11843. The tissuelyser ii is an integral part of qiagens complete solution for sample management from sample collection to purification and analysis of dna, rna, and protein. Rna concentration and rna integrity were assessed using the agilent 4200 tapestation agilent, santa clara, ca. Preparation of cytosolic and microsomal fractions was performed as described previously.
Does anyone have experience with tissue lyser from qiagen. See the tissuelyser lt handbook or the tissuelyser handbook. Safety information 6 tissuelyser ii user manual 042010 with accessories approved by qiagen see section 3. I tried a tissue lyser from qiagen to disrupt some mice aortas in order to extract protein. Total rna extraction from tissues for microrna and target. I used it at maximal speed, as these tissue are really elastic and difficult to disrupt. May 2016 tissuelyser ii user manual gtac home page. The tissuelyser ii complements qiagen automation for highthroughput sample preparation and analysis see table qiagen highthroughput automation, such as the qiasymphony sp. Amplifications were carried out on an eppendorf pcr cycler using the following pro file. Qiagen rotorgene q pcr cycler 5 plex, qiagility prep. The tissueruptor and tissuelyser ii are integral parts of qiagens complete solution for tissue management from sample collection to. Table 1 comparison of the five microbial dna extraction methods used in this study. Carbonyl reduction of nnk by recombinant human lung.
Integrated in qiagen tissue management workflow tissueruptor and tissuelyser ii disruption systems for low to highthroughput disruption of biological samples qiagen provides 2 systems for fast, effective sample disruption. Qiagen contact details, which may be required to place orders or to request technical support, are also displayed. Qiaex ii handbook qiaex ii agarose gel extraction protocol. Qiagen offers advanced and innovative solutions for all of your assay and sample equipment needs. Evaluating variation in human gut microbiota profiles due.
The tissuelyser ii is wellsuited for highthroughput disruption of human, animal, and plant tissues, bacteria, and yeast. The qiagen knowledge base is an industryleading database com. Qiaquick gel extraction kit protocol using a microcentrifuge this protocol is designed to extract and purify dna of 70 bp to 10 kb from standard or lowmelt agarose gels in tae or tbe buffer. It has been stored under unfavorable conditions for a prolonged period. Tissuelyser adapter set 2 x 96 from qiagen selectscience. Comparison of dna extraction methods for human gut. Tissueruptor tissuelyser lt tissuelyser ii throughput samplesrun 1 up to 12 up to 48 or 192 disruption technology rotorstator bead mill bead mill disruption time 30 seconds 40 seconds to 5 minutes 20 seconds to 5 minutes table 1. These are available online in convenient and compact pdf. Frontiers all in one high quality genomic dna and total. Comparison of dna extraction methods for human gut microbial. Lung 60 50 40 30 20 10 0 tissuelyser lt tissuelyser ii liver heart brain skin 400 300 200 100 0 heart brainlung tissuelyser lt tissuelyser ii figure 6. File save as menu item or clicking the save file toolbar icon. Tissuelyser ii user manual 052016 7 it or the line power cord appears to be damaged.
Qiagen multiplex pcr kit, x 50l rxns, genomic dna and cdna sample, pcr amplification reaction, 5 3 exonuclease enzyme activity, easytouse and costeffective, for highly specific and sensitive multiplex pcr without optimization requirements, includes 2x qiagen. Qiagen is the leading provider of innovative sample and assay technologies, enabling. In preparation for rna extraction 510 mg of tissue weights differed by tissue but were consistent for all kits was added to 700. Evaluation of parameters affecting switchgrass tissue culture. Various rat tissues were disrupted using the tissuelyser lt or.
Place a qiaquick spin column in a provided 2 ml collection tube or into a vacuum manifold. Page 156 use the file open menu item or open file toolbar item to open an existing run file. Switchgrass panicum virgatum, a robust perennial c4type grass, has been evaluated and designated as a model bioenergy crop by the u. Qiagen shall not be held responsible for any injuries or damages resulting from nonobservance of the instructions and safety information in this user manual. Tissue homogenization was performed with a qiagen tissuelyser ii in 2 ml tubes with a 5mm bead at 22 hz for 2 min. There are minor cosmetic scratches on the top plate, but nothing serious.
Bioanalyzer at the vanderbilt genome sciences resource. Microbiome dysbiosis is associated with disease duration and. Click no to discard the changes and proceed with the requested operation. All microsomes were washed once with trisbuffered saline ph 7. This easytouse instrument automates purification of dna, rna, and protein from 196 samples.
Evaluation of parameters affecting switchgrass tissue. Doubleclick or choose open to open the chosen run file. Rna isolation with trizol invitrogen and qiagen rnaeasy. Used qiagen mm300 tissuelyser shaker mixer grinder. The tissuelyser lt is compact so you can keep it nearby for easy access. Qiagen offers adapter sets for 2 x 96 collection microtubes 1. Does anyone use the tissuelyser ii qiagen equipment to. About qiagility displays information about the qiagility, including the version number, serial number and firmware number. The samples were spun down 14,000 rpm, 4 c, 10min, the pulverized aqueous sample 900l transferred to a fresh micro. Jun 02, 2014 add the appropriate volume of buffer rlt see table 8, and homogenize by passing the lysate at least 5 times through a blunt 20gauge needle fitted to an rnasefree syringe. Equipnet is constantly receiving used qiagen equipment through our exclusive contracts with our clients. Tissuelyser ii from qiagen, one stainless bead per 2ml tube used, homogenization at a frequency 30s for 3x1 min.
Qiagen disruption systems for a range of throughputs. For optimal operation, the tissuelyser must always be balanced. It has been subjected to severe transport stresses. The qiagen tissuelyser lt sample disruptor provides quick and efficient disruption in up to 12 samples. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease. Disruption and homogenization using the tissuelyser lt or tissuelyser ii.
Laboratory equipment qiagen tissuelyser ii user manual. Total rna was extracted and cdna was generated 82 using a high capacity cdna kit life technologies. This german made shaker appears in good condition without cracks in the plastic cover or other damage to the housing or workings. Add 1 gel volume isopropanol to the sample and mix. A detailed investigation of the porcine skin and nose microbiome using universal and staphylococcus. This kit can also be used for dna cleanup from enzymatic reactions see page 8. Sequencebased analysis of the bacterial and fungal. Tissuelyser ii laboratory equipment pdf manual download. Qiagen tissuelyser ii sample disruption homogenizer system see details.
Used qiagen mm300 tissuelyser shaker mixer grinder agitator. For efficient disruption of up to 48 or 192 samples in parallel on the tissuelyser ii. Extraction was optimised with a 10 min 70 c incubation quigley et al. Tissuelyser ii user manual en print bookmark share pdf 2185kb english. We used 2030mg of tissue material in 600ll of buffer rlt plus with 6ll b.
Tissueruptor and tissuelyser ii disruption systems biomarker. Our inventory of used qiagen equipment includes various detectors, dna synthesizers, sample equipment, liquid handlers, and much more. Qiagen tissuelyser lt sample homogenizer system see details. Spin down at 10,000g to remove remaining solid material, use supernatant for further processing. Mar 16, 2018 in preparation for rna extraction 510 mg of tissue weights differed by tissue but were consistent for all kits was added to 700. The tissuelyser ii is an integral part of qiagen s complete solution for sample management from sample collection to purification and analysis of dna, rna, and protein. The powerfood microbial dna isolation kit was then used as per the manufacturers instructions. If the sample is not fully homogenized, repeat with decreased time and decreased frequency 1 minute, 20 hz for no more than 2 additional minutes. View and download qiagen tissuelyser ii user manual online.
Print bookmark share more for highthroughput disruption of biological samples. Fix the adapter sets into the clamps arms of the tissuelyser. This small bead mill has high throughput with highspeed shaking for the disruption or homogenization of samples in 2 ml microcentrifuge tubes with stainless steel or glass beads. Using the appropriate adapter set, up to 48 or 192 samples can be processed at the same time.