Contents 1 2 2 3 4 5 6 8 10 11 12 13 15 Introduction Advantages Implementation on robotic workstations Product selection guide Smarter DNA sample preparation DNA isolation procedures Invisorb® Plant DNA Kits InviMag® Plant DNA Kits Smarter RNA sample preparation RNA isolation procedures InviTrap® Plant RNA Kits References about STRATEC Molecular Smarter Nucleic Acid Sample Preparation for Plant Applications Patented complete solutions for customer applications Contact information STRATEC Molecular GmbH Robert-Rössle-Str. 10 D-13125 Berlin Germany Phone: +49-30-9489 2901/2910 Fax: +49-30-9489 3795 Email: [email protected] www.invitek.de 6I7d06/05/2011 Sample preparation kits for the isolation of DNA and RNA from: fresh, frozen or dried plant material food of plant origin Introduction Plant genotyping, DNA fingerprinting of plants, microsatellite analysis or single nucleotide polymorphism (SNP) marker analysis are technologies that have matured and are poised for widespread practical applications. Plant genotype analysis can be used for the identification of plants in commerce, plant breeding and research. Further it allows the analysis of wild plant populations, germplasm collections and plant protection. In food diagnostics GMOs have to be identified in soy or corn compounds. The presence of GMOs can affect the use and value of products of an entire lot. For these purposes STRATEC Molecular offers high-performance DNA and RNA isolation kits for a wide variety of plant material incl. food from plant origin. Under the trademarks Invisorb®, InviTrap® and InviMag® STRATEC Molecular offers a growing panel of DNA and RNA isolation kits for plant materials starting from single sample preparations up to 96 well isolation systems using different extraction principles like binding to membranes or to magnetic particles. 1 Advantages o pure, ready-to-use DNA/RNA for enhanced performance in sensitive downstream applications o various isolation systems available to suit your special needs o cross contamination free processes o ready-made and customized purification protocols Implementation on robotic workstations The adaptation of well established STRATEC Molecular kits to common liquid handling workstations allows for a simple and automated isolation of nucleic acids. The procedures are designed to avoid sample-to-sample cross contamination. The InviMag® DNA Plant Kits for medium and high throughput applications using magnetic particles are designed for use on the KingFisher mL, KingFisher 96 and KingFisher Flex platforms. o special customer tailored solutions are also available on request o programs can simply be downloaded from www.invitek.de o for adaptation to robotic platforms please contact STRATEC Molecular: +49 30 9489 2901 or [email protected] 2 Product selection guide Format Starting material Article number Product name Package size DNA purification Single tube, membrane adsorption based HTS, membrane adsorption based Single tube, magnetic bead based HTS semiautomated, magnetic bead based 100 - 300 mg fresh or frozen and up to ® Invisorb Spin Plant Mini Kit 60 mg dried plant material 1037100200 1037100300 50 preps 250 preps up to 500 mg fresh or frozen plant material Invisorb Spin Plant Midi Kit ® 1037110200 1037110300 25 preps 50 preps up to 100 mg fresh or frozen fresh, processed or preserved food sample of vegetable origin Invisorb Spin Food Kit II ® 1036110200 1036110300 50 preps 250 preps up to 50 mg fresh or frozen plant material Invisorb DNA Plant HTS 96 Kit/ C ® 7037300200 2 x 96 preps 7037300300 4 x 96 preps 7037300400 24 x96 preps up to 50 mg fresh or frozen plant material Invisorb DNA Plant HTS 96 Kit/ V up to 100 mg fresh or frozen plant material (96 well; using a centrifuge) ® (96 well; using a vacuum manifold) ® InviMag Plant DNA Mini Kit (for manual use on a magnetic rack) ® up to 100 mg fresh or frozen plant material InviMag Plant DNA Mini Kit/ KFmL up to 100 mg fresh or frozen plant material InviMag Plant DNA Mini Kit/ KF96 (automated; for use on KingFisher mL) 7037310200 2 x 96 preps 7037310300 4 x 96 preps 7037310400 24 x96 preps 1437100200 1437100300 50 preps 250 preps 2437110200 2437110300 2437110400 75 preps 150 preps 300 preps ® (automated; for use on KingFisher 96 and KingFisher Flex) 7437300100 1 x 96 preps 7437300200 5 x 96 preps RNA purification Single tube, membrane adsorption based HTS, membrane adsorption based 7 up to 100 mg or 1x10 cells of plant material or filamentous fungi ® 1064100200 1064100300 ® 7064300200 2 x 96 preps 7064300300 4 x 96 preps 7064300400 24 x96 preps InviTrap Spin Plant RNA Mini Kit 25 preps 50 preps up to 50 mg fresh or frozen plant material InviTrap RNA Plant HTS 96 Kit/ C up to 50 mg fresh or frozen plant material InviTrap RNA Plant HTS 96 Kit/ V up to 50 mg fresh or frozen plant material InviTrap RNA Plant HTS 96 Kit/ R for parallel purification of genomic DNA from the same sample add-on module for InviTrap RNA Plant 7064000200 2 x 96 preps HTS 96 Kit 7064000300 4 x 96 preps (96 well format; for centrifuge & robot) 7064000400 24 x96 preps (96 well; using a centrifuge) ® (96 well; using a vacuum manifold) ® (automated 96 well; using a robot) ® 3 7064310200 2 x 96 preps 7064310300 4 x 96 preps 7064310400 24 x96 preps 7164300200 2 x 96 preps 7164300300 4 x 96 preps 7164300400 24 x96 preps Smarter DNA sample preparation Superior non-chaotropic chemistry – low salt DNA binding o Improve Effectiveness - More intact DNA no DNA degradation by using a gentle, low-salt (non-chaotropic) buffer system o Improve Effectiveness - High yields and purity pure DNA free of polysaccharides and other secondary metabolites o Maximize Capabilities - For various plant materials leaves, roots, fruits, flowers, wood, oily seeds o Maximize Capabilities - Downstream applications suitable for PCR applications, restriction analysis, real-time PCR, cloning, genotyping etc. Optimized protocols The homogenized sample is lysed in an optimized lysis buffer and proteins are degraded during the lysis with Proteinase K. Using membranes in spin columns or 96 well filter plates the cell debris is removed via prefiltration. In the magnetic bead based procedure this step is not necessary. The DNA binds to the filter membrane or magnetic particles, followed by washing steps and the final elution. The purified genomic DNA is of high quality and can be used for a broad panel of subsequent downstream applications: o o o o o o o PCR real-time PCR RFLP analysis RAPD analysis AFLP analysis Southern Blotting GMO analysis 4 DNA isolation procedures spin column - membrane 96 well - membrane Lysis of homogenized starting material Lysis of homogenized starting material Precleaning Precleaning Adjustment of binding conditions Adjustment of binding conditions Waste Waste magnetic beads Lysis of homogenized starting material Addition of ® InviMag beads and Binding Solution to the lysate DNA binds to magnetic particles Magnetic separation Binding of DNA Washing of the particle fixed DNA Binding of DNA Washing Removal of ethanol Washing Removal of ethanol Elution of pure DNA Elution of pure DNA Magnetic separation Elution of DNA Magnetic separation Elution of pure DNA 5 Invisorb® Plant DNA Kits The Invisorb® Spin Plant Mini Kit is an effective solution for isolation of high-quality total cellular DNA from a wide variety of plant species and tissue types. Up to 100 mg of fresh or frozen material can be processed in less than one hour. The optimized procedure incorporates the Pre-filtration spin column, a unique filtration and homogenization column that efficiently removes cell debris and improves sample handling following lysis. Up to 35 µg pure DNA can be obtained, free of polysaccharides and other secondary metabolites. For larger sample volumes (up to 500 mg plant material) the Invisorb® Spin Plant Midi Kit can be used. Fig. 1 RAPD analysis of Brassica and Raphanus species DNA was isolated from leaves of Raphanus sativus, Brassica oleracea and hybrids of both species using the Invisorb® Spin Plant Mini Kit. DNA (20 ng) was amplified with the operon-decamer primer OPC-10 and separated on a 4% polyacrylamide gel. lane 1 lane 2 lane 3 lane 4 lane 5 lane 6 lane 7 lane 8 1 2 3 4 5 6 7 8 100 bp ladder (GIBCO BRL) Raphanus sativus plant I Raphanus sativus plant II Brassica oleracea plant I Brassica oleracea plant II hybrid I Raphanus x Brassica hybrid II Raphanus x Brassica 100 bp ladder (GIBCO BRL) (Data kindly provided by Dr. Budahn, Institute for Breeding Methods of vegetables; Federal Centre for Breeding Research on Cultivated Plants; Quedlinburg) Fig. 2 AFLP analysis of Chlorella vulgaris DNA was isolated from different strains of Chlorella vulgaris using the Invisorb® Spin Plant Mini Kit. Isolated DNA (100 ng) was digested with EcoR1 and Msel followed by selective amplification with EcoRI-C/MseI-C primers. The AFLP-analysis was carried out on ABI PRISM 3100 Genetic Analyzer. (Data kindly provided from Frau J. Müller, Albrecht-von-Haller-Institute for plant science, University of Göttingen) 6 Invisorb® Plant DNA Kits The Invisorb® Spin Food Kit II simplifies DNA purification from food of plant origin. A wide variety of starting materials can be processed, e.g. fresh, dried, preserved, cooked, or frozen vegetable tissues and processed foods like flour, tofu, corn chips, bread, and cookies. The procedure yields high-quality DNA that is ready to use in downstream applications, such as sensitive DNA-based detection of genetically modified crops (GMOs) in various materials (e.g. soy and corn). 1 A A 2 3 4 5 1 2 3 5 6 7 8 B Lane 1/ 8 Lane 2 100 bp ladder (NEB) soy, 35 S1/2 (CaMV-promotor, 195 bp), contains 1.5% GVO-amount according the results of the interlaboratory test maize, 35 S1/2 (CaMV-promotor, 195 bp) mixed fodder, 35 S1/2 (CaMV-promotor, 195 bp) 100 bp ladder (NEB) soy, soy detection GM 03/04 (lectin-gene, 118 bp) maize, specific GVO-detection T25 (CaMV- pat-gene, 209 bp (contains 0,1 % T25- amount according the results of the interlaboratory test) Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Fig. 3 4 Isolation of genomic DNA from food and feed DNA was isolated from soy and maize samples of an interlaboratory test and from an animal feed sample of different plants with the Invisorb® Spin Food Kit II. 5 µl of the eluted DNA were analyzed on a 2% agarose gel (figure A). The isolated DNA (A) was amplified. The PCR analysis was performed with specific primer according § 35 LMBG method. 20 µl of the amplified product were analyzed on a 2 % agarose gel as shown in figure B. (Data kindly provided by Mrs. Häger, AG Dr. Baier, JenaGen GmbH, Jena) The Invisorb® DNA Plant HTS 96 Kit/ C is designed for convenient DNA purification from up to 50 mg of a wide variety of plant species in 96 well format using filter plates and a centrifuge. A vacuum-driven process is available on request. There is also the possibility to automate this process. B A Fig. 4 DNA isolation from Arabidopsis thaliana and subsequent PCR amplification DNA was isolated from single leaves from Arabidopsis thaliana using the Invisorb® DNA Plant HTS 96 Kit/ C on a deep well rotor centrifuge. The leaves were homogenized before lysis by using a Retsch Mixer mill under liquid nitrogen. The DNA was eluted in 60 µl and 5 µl were used in a subsequent PCR reaction. 10 µl of the eluted DNA were analyzed on a agarose gel (figure A) and 10 µl of the PCR products were analyzed on a 2% agarose gel (figure B). 7 A B InviMag® Plant DNA Kits The InviMag® Plant DNA Kits allow for rapid and economical purification of genomic DNA from up to 100 mg of fresh, frozen or dried plant material using magnetic beads. The InviMag® Plant DNA Mini Kit is designed for manual use on the InviMag® Rack. The InviMag® Plant DNA Kits/ KF are designed for automated preparation on the KingFisher workstations. 1 2 3 4 5 6 7 8 Fig. 5 Analysis of plant DNA DNA from each 60 mg of Allium cepaand and from 60 mg Hedera helix was isolated using the InviMagPlant DNA Mini Kit/ KFmL on KingFisher mL and 10 µl of the eluate were analyzed after an RNAse A treatment on a 0.8 % agarose gel. Lane 1, 8 Lane 2 - 4 Lane 4 - 7 10 kb ladder DNA from Allium cepaand DNA from Hedera helix Fig. 6 Analysis of DNA yield from different plant species DNA from 60 mg of cress, leaves of clover, or maize, maize seeds and parsley was isolated using the InviMagPlant DNA Mini Kit/ KF 96 on the KingFisher 96 and the yield was detected using a spectrophotometer for the different species after an RNase A treatment. 8 InviMag® Plant DNA Kits Identification of GMOs in corn compounds Examples of GMOs are diverse, and include transgenic experimental animals such as mice, several fish species, transgenic plants or various microscopic organisms altered for the purposes of genetic research or for the production of pharmaceuticals. The presence of GMOs can affect the use and value of products in an entire lot. Genomic DNA from Corn positive control standard sample Red curve – positive control Light green curves – samples Dark green curves – standard GMO determination positive control sample standard Red curve – positive control Light blue curves – samples Dark blue curves – standard Fig. 7 DNA was isolated from 60 mg hackled corn compounds including defined amounts of GMO material, using the InviMag® Plant DNA Mini Kit/ KFmL. The DNA was eluted in 100 µl Elution Buffer. 5 µl of the eluted DNA were used for the real-time amplification of genomic DNA and GMO-DNA, to determine the copy number. By measurement of genomic and GMO copies the relation between the two genetic components was calculated. It was compared with the known data of the materials. Percentage of GMO copies to DNA copies Corn compounds A Genomic DNA copies 10.715 GMO DNA copies 0 Ratio Genomic DNA: GMO DNA 0 Theoretical amount of GMO copies 0 0.02 % B 15.009 5 0.03 % C 11.556 23 0.2 % 0.3 % D 9.422 70 0.74 % 1.0 % E 10.205 527 5.2 % 8.0 % (Data kindly provided from Dr. St. Mergemeier, Congen Biotechnologie GmbH) 9 Smarter RNA sample preparation InviTrap® - pure RNA without DNase The unique feature of InviTrap® products is the specific removal of DNA from RNA sample preparations. Efficient RNA purification from plants o Improve Efficiency - Pure RNA without DNase digestion selective genomic DNA removal during lysis step o Improve Effectiveness - High yields up to 100 µg pure RNA from 100 mg plant material or 107 plant cells o Maximize Capabilities - Optimal processing of various samples two sample-specific lysis buffers included - contaminants, such as polysaccharides, or polyphenols are removed o Maximize Capabilities - Versatile use incl. simultaneous isolation of RNA and proteins or RNA and DNA Optimized protocols The homogenized sample is lysed in an optimized and RNase inactivating buffer. To satisfy the variable composition of plants two sample-specific lysis buffers are included. Contaminants such as polysaccharides and polyphenols are efficiently removed. During lysis the genomic DNA binds to the surface of specific carrier particles. Digestion with DNase is not necessary. After removal of the cell debris and the DNA-binding carrier the RNA binding conditions are adjusted. Total RNA binds to the membrane of the spin column or 96 well plate followed by washing steps, leaving pure RNA to be eluted in RNase free water. The isolated RNA is ready-to-use for subsequent downstream applications, e.g.: o RT-PCR o real-time RT-PCR o Northern Blotting 10 RNA isolation procedures spin column - membrane 96 well - membrane Lysis of homogenized starting material Lysis of homogenized starting material Adjustment of DNA binding conditions Waste Selective removal of genomic DNA Adjustment of DNA binding conditions Adjustment of RNA binding conditions Selective removal of genomic DNA Waste Adjustment of RNA binding conditions Binding of RNA Binding of RNA Washing steps Elution of pure RNA Washing steps Elution of pure RNA 11 InviTrap® Plant RNA Kits The InviTrap® Spin Plant RNA Mini Kit is the ideal tool for fast and easy isolation of total RNA from a wide variety of plant samples and filamentous fungi. Pure RNA can be isolated from up to 100 mg of plant material or 107 plant or fungal cells in less than 30 min. The genomic DNA is removed without an enzymatic digestion step. RNases are inactivated to prevent RNA degradation. Furthermore the InviTrap® RNA kits feature capped spin columns for safe and reliable RNA purification. Fig. 8 High-quality total RNA from Arabidopsis thalianan Total RNA was isolated from Arabidopsis thalianan (leaves) using the InviTrap® Spin Plant RNA Mini Kit. RNA was separated on a denaturating agarose gel. (Data kindly provided from Ms. Weißleder, Leibniz Institute of Plant Genetics and Crop Plant Research, Gatersleben, Germany) Fig. 9 Excellent RNA performance in downstream RT-PCR assay Total RNA was isolated from 100 mg of fresh leaves or young shoots of orange, mandarin and grapefruit plants using the InviTrap® Spin Plant RNA Mini Kit. RNA was eluted in 50 µl of elution buffer and amplified in a RT-PCR assay using viroid specific CVd-IIb primers. Lane M, 50 bp DNA ladder; -, water; H, healthy citrus; +, positive control; 1-4, tested citrus samples (Survey and molecular detection of two citrus viroids affecting commercial citrus orchards in the Northern part of Sudan. Mohamed Yousif Adam Abubaker and Siddig Mohamed Elhassan. Agric. Biol. J. N. Am., 2010, 1(5): 930-937) The InviTrap® RNA Plant HTS 96 Kit/ C simplifies RNA isolation from 96 plant samples in parallel using a filter plate in a centrifuge. The genomic DNA is not digested with DNase but removed using a DNA-binding carrier. Therefore simultaneous DNA isolation is possible via an additional module. A vacuum-driven process is available on request. There is also the possibility to automate this process. Fig. 10 RNA isolation from transgenic Populus tremula x tremuloides RNA was isolated from young leaves of heat-shock inducible transgenic lines of hybid aspen (Populus tremula x tremuloides) (1-6) and from more difficult tissue-culuture grown stem (8), apex (7) and greenhouse leaves (9) after disruption of the plant material in a bead mill using the InviTrap® RNA Plant HTS 96 Kit/ C. The figure shows a representative selection of RNA samples, the sharp ribosomal RNA banding pattern shows that the kit produces high quality intact RNA from young leaves, and also from more difficult tissues such as steam. (Data kindly provided from A. Karlberg, Umea Plant Science centre, Swedish University of Agricultural Sciences, Umea, Sweden) 12 References Plant DNA Phylogeny of the leafy liverwort Ptilidium: cryptic speciation and shared haplotypes between the Northern and Southern Hemispheres. Kreier HP, Feldberg K, Mahr F, Bombosch A, Schmidt AR, Zhu RL, von Konrat M, Shaw B, Shaw AJ, Heinrichs J. Mol Phylogenet Evol. 2010 Dec;57(3):1260-7 One species or at least eight? Delimitation and distribution of Frullania tamarisci (L.) Dumort. s. l. (Jungermanniopsida, Porellales) inferred from nuclear and chloroplast DNA markers. Heinrichs J, Hentschel J, Bombosch A, Fiebig A, Reise J, Edelmann M, Kreier HP, Schäfer-Verwimp A, Caspari S, Schmidt AR, Zhu RL, von Konrat M, Shaw B, Shaw AJ. Mol Phylogenet Evol. 2010 Sep;56(3):1105-14 Mycotoxic nephropathy in Bulgarian pigs and chickens: complex aetiology and similarity to Balkan endemic nephropathy Stoev SD, Dutton MF, Njobeh PB, Mosonik JS, Steenkamp PA. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2010 Jan;27(1):72-88 Diversification and Taxonomy of the Liverwort Jubula Dumort. (Jungermanniopsida: Porellales) Inferred from Nuclear and Chloroplast DNA Sequences Pätsch, Ricarda; Hentschel, Jörn; Linares-Palomino, Reynaldo; Zhu, Rui-Liang; Heinrichs, Jochen Systematic Botany (2010), 35(1): pp. 6–12 In vitro vs in silico detected SNPs for the development of a genotyping array: what can we learn from a non-model species? Lepoittevin C, Frigerio JM, Garnier-Géré P, Salin F, Cervera MT, Vornam B, Harvengt L, Plomion C. PLoS One. 2010 Jun 9;5(6):e11034. High level protein expression in plants through the use of a novel autonomously replicating geminivirus shuttle vector. Regnard GL, Halley-Stott RP, Tanzer FL, Hitzeroth II, Rybicki EP. Plant Biotechnol J. 2010 Jan;8(1):38-46 Genome size correlates with growth form, habitat and phylogeny in the Andean genus Lasiocephalus (Asteraceae). Eva Dušková, Filip Kolář, Petr Sklenář, Jana Rauchová, Magdalena Kubešová, Tomáš Fér, Jan Suda & Karol Marhold; Preslia 82: 127–148, 2010 Studies on Lophocoleaceae. XIX. The systematic identity of Cyanolophocolea R.M. Schust., an intriguing liverwort from New Zealand and Australia, based on morphological and molecular evidence. John J. Engel and Xiaolan He The Bryologist 113(1), pp. 149–163 Natural hybridization in tropical spikerushes of Eleocharis subgenus Limnochloa (Cyperaceae): Evidence from morphology and DNA markers Jan Kosnar, Jirí Kosnar, Miroslava Herbstová, Petr Macek, Eliska Rejmánková, and Milan Stech Am. J. Botany, Jul 2010; 97: 1229 - 1240. Reexamination of the genus Pterocladiella (Gelidiaceae, Rhodophyta) in Korea based on morphology and rbcL sequences. Sung Min Boo, Su Yeon Kim, In Sun Hong and Il Kiwang Algae 2010, 25(1): 1-9 First Report of Aster Yellows Phytoplasma in Grapevines in South Africa. M. Engelbrecht, J. Joubert, J. T. Burger Plant Disease; March 2010, Volume 94, Number 3; Page 373 Hylodesmus singaporensis gen. et sp. nov., a new autosporic subaerial green alga (Scenedesmaceae, Chlorophyta) from Singapore. Marek Eliás, Yvonne Nemcová, Pavel Skaloud, Jirí Neustupa, Veronika Kaufnerová, and Lenka Sejnohová Int J Syst Evol Microbiol, May 2010; 60: 1224 - 1235 Isolation of microsatellite markers for the common Mediterranean shrub Myrtus communis (Myrtaceae). Rafael G. Albaladejo, Federico Sebastiani, Santiago C. González-Martínez, Juan P. González-Varo, Giovanni G. Vendramin, and Abelardo Aparicio Am. J. Botany, May 2010; 97: e23 - e25 Plant RNA Recognition of A. thaliana centromeres by heterologous CENH3 requires high similarity to the endogenous protein. Moraes IC, Lermontova I, Schubert I. Plant Mol Biol. 2011 Feb;75(3):253-61 Over-expression of an FT-homologous gene of apple induces early flowering in annual and perennial plants. Tränkner C, Lehmann S, Hoenicka H, Hanke MV, Fladung M, Lenhardt D, Dunemann F, Gau A, Schlangen K, Malnoy M, Flachowsky H. Planta. 2010 Nov;232(6):1309-24 RLK7, a leucine-rich repeat receptor-like kinase, is required for proper germination speed and tolerance to oxidative stress in Arabidopsis thaliana. Pitorre D, Llauro C, Jobet E, Guilleminot J, Brizard JP, Delseny M, Lasserre E. Planta. 2010 Nov;232(6):1339-53 Kinome profiling reveals an interaction between jasmonate, salicylate and light control of hyponastic petiole growth in Arabidopsis thaliana. Ritsema T, van Zanten M, Leon-Reyes A, Voesenek LA, Millenaar FF, Pieterse CM, Peeters AJ. PLoS One. 2010 Dec 8;5(12):e14255. Expression divergence of the AGL6 MADS domain transcription factor lineage after a core eudicot duplication suggests functional diversification. Viaene T, Vekemans D, Becker A, Melzer S, Geuten K. BMC Plant Biol. 2010 Jul 15;10:148 The proteome map of spinach leaf peroxisomes indicates partial compartmentalization of phylloquinone (vitamin K1) biosynthesis in plant peroxisomes Lavanya Babujee, Virginie Wurtz, Changle Ma, Franziska Lueder, Pradeep Soni, Alain van Dorsselaer, and Sigrun Reumann J. Exp. Bot., Mar 2010; 61: 1441 - 1453. 13 References Two novel proteins expressed by the venom glands of Apis mellifera and Nasonia vitripennis share an ancient C1q-like domain. de Graaf DC, Brunain M, Scharlaken B, Peiren N, Devreese B, Ebo DG, Stevens WJ, Desjardins CA, Werren JH, Jacobs FJ. Insect Mol Biol. 2010 Feb;19 Suppl 1:1-10. A Study of Gene Expression in the Nematode Resistant Wild Peanut Relative, Arachis stenosperma, in Response to Challenge with Meloidogyne arenaria. Patricia Messenberg Guimarães, Ana Cristina Miranda Brasileiro, Karina Proite, Ana Claudia Guerra de Araújo, Soraya Cristina Macedo Leal-Bertioli, Aline PicTaylor, Felipe Rodrigues da Silva, Carolina Vianna Morgante, Simone da Graça Ribeiro and David John Bertioli Tropical Plant Biol., DOI 10.1007/s12042-010-9056-z Survey and molecular detection of two citrus viroids affecting commercial citrus orchards in the Northern part of Sudan. Mohamed Yousif Adam Abubaker and Siddig Mohamed Elhassan Agric. Biol. J. N. Am., 2010, 1(5): 930-937 Identification of host genes potentially implicated in the Malus pumila and ‘Candidatus Phytoplasma mali’ interactions. M. Aldaghi, S. Massart, A. Bertaccini, P. Lepoivre 21st International Conference on Virus and other Graft Transmissible Diseases of Fruit Crops; Julius-Kühn-Archiv, 427, 2010 Identification and evaluation of new reference genes in Gossypium hirsutum for accurate normalization of real-time quantitative RT-PCR data. Artico S, Nardeli SM, Brilhante O, Grossi-de-Sa MF, Alves-Ferreira M. BMC Plant Biol. 2010 Mar 21;10:49. The Unique Biosynthetic Route from Lupinus β-Conglutin Gene to Blad. Monteiro S, Freitas R, Rajasekhar BT, Teixeira AR, Ferreira RB (2010) PLoS ONE 5(1): e8542 Abscisic Acid-Induced Resistance against the Brown Spot Pathogen Cochliobolus miyabeanus in Rice Involves MAP Kinase-Mediated Repression of Ethylene Signaling. David De Vleesschauwer, Yinong Yang, Casiana Vera Cruz, and Monica Höfte Plant Physiology, Apr 2010; 152: 2036 - 2052. 14 About STRATEC Molecular STRATEC Molecular – part of the STRATEC group since 2009 – is a globally active provider of innovative system solutions for nucleic acid sample collection, stabilization, and both manual and automated purification from any sample type. Since 1992 the company is internationally respected for its outstanding and high performance technology platforms and offers a broad spectrum of superior products for molecular diagnostics, drug discovery and Life Science research. As an EN ISO 13485:2003 + AC 2007 and EN ISO 9001:2008 certified company all STRATEC Molecular products are subject to extensive quality control. In compliance with Directive 98/79/EC (IVDD) many STRATEC Molecular products are CE-certified. STRATEC Molecular guarantees the correct function of all products and highest quality support by first-rate service. About the STRATEC group The STRATEC group consists of the publicly listed parent company STRATEC Biomedical AG and of subsidiaries and second-tier subsidiaries in Germany, the USA, the UK, Switzerland and Romania. The STRATEC Biomedical AG (http://www.stratec.com) designs and manufactures fully automated systems for its partners in the fields of clinical diagnostics and biotechnology. Core technologies ® InviTrap - Selective removal of DNA Non-chaotropic chemistry - shorter protocols through reduced salt concentrations higher yields from complex/precious samples more intact chromosomal DNA - - ® ® InviMag - Magnetic beads MSB - Minimal salt binding - - the fastest way to purify DNA fragments excellent purity without washing - RNAsure - Viral RNA protection - Extraction Tube provides all lysis components, Carrier RNA and standards stabilized at RT safer sample handling due to reduced hands-on steps - ® PSP - DNA sample stabilization - manual and automated DNA and RNA purification ® ® RTP - Ready to prep - highly purified RNA for better RT-PCR results no DNase digestion required stabilization of host/pathogen DNA at RT in stool, saliva or swab samples preservation of bacterial titers at the time of sampling 15 immediate lysis and viral RNA stabilization in biological samples RNA resistant to degradation for up to 6 month at RT
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