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Nucleic acid extraction and purification technologies For research use only. Not for use in diagnostic procedures Extraction and DNA extraction purification solutions chemistries from LGC, Biosearch QuickExtract Technologies For DNA screening and genotyping-type applications, PCR-grade genomic DNA is sufficient, which can be rapidly and efficiently Biosearch Technologies is unique in the nucleic DNA extraction chemistries extracted using QuickExtract™ DNA Extraction acid extraction and purification marketplace, 3 QuickExtract Solution. In 3-8 minutes, and using only a single- in that we are both a service provider, and tube protocol, PCR-grade genomic DNA from a developer of proprietary chemistries and DNA purification chemistries almost any sample type can be extracted, without instrumentation. We develop chemistries and 4-6 PCR-grade purifications – Kleargene the use of centrifuges or spin-columns (Figure 1). instrumentation for customers to use in their NGS-grade purifications – sbeadex (plant, Add samples own laboratories, and it is these same products livestock, blood) QuickExtract Solution has been used to which we use in each of our service laboratories. extract DNA from samples such as hair This allows Biosearch Technologies to be highly 7 DNA/RNA/total nucleic acid follicles, quill-end cells of feathers, tissue- flexible, and offer individual solutions for each of purification chemistries QuickExtract culture cells, buccal cells, zebrafish organs our customers. MasterPure solution and scales, and mouse tail snips. The To provide the most efficient and effective Heat at 65 ˚C for 6 minutes extracted DNA is suitable for PCR analyses extraction or purification method, we offer not 8 Automation and 98 ˚C for 2 minutes (Figure 2), such as genomic, transgenic, only a range of different technologies but also the or viral DNA screening in animals, or for capability to develop tailor-made kit systems for all Purification services genetic or environmental research and applications and downstream requirements. 9 DNA purification services PCR-ready screening in humans and other organisms. Figure 1: Procedure for obtaining DNA RNA purification services PCR-ready DNA using QuickExtract Our range of technologies allows delivery of DNA Extraction Solution. optimised extractions and purifications from an 10 Overview unrivalled array of matrices including: The range of QuickExtract Extraction Solutions for • Plant material (including leaves, seeds, and 11 Order information additional sample types includes: other plant matrices) • QuickExtract DNA Extraction Solution • Livestock samples (including blood, animal • QuickExtract FFPE DNA Extraction Solution tissue, hair follicle, buccal swabs and saliva) • QuickExtract Plant DNA Extraction Solution • Human samples (including blood, tissue and • QuickExtract RNA Extraction Solution forensic samples) • Plasmid preparations • Bacteria, prokaryotes and other Figure 2: Samples extracted with QuickExtract DNA Extraction Solution. microorganisms PCR was performed using primers to amplify the regions indicated: Lanes 1-3, human β-globin; lane 4, transgenic mouse GAPDH; lane 5, E. coli 16S ribosomal RNA gene; lane 6, transgenic SV40 T antigen. 2 3 need of a drying stage, and minimising any sbeadex plant DNA purification contaminating ethanol carryover into the final eluate. The switch between binding mechanisms Plant samples are often challenging due to also allows for greater purity of nucleic acids. secondary metabolites, high levels of sugars and chemistries The advantages of sbeadex include: polyphenols, or robust cell walls. sbeadex plant protocols for leaf and/or seed material from a broad Kleargene has been highly optimised for • High-quality DNA: suitable for all genomic- range of plant species have been validated and PCR-grade DNA automation with both 96-well formats (plant and applications (including NGS) can be automated on all open platforms, including tissue) and 384-well formats (plant), achieving • Compatible with Biosearch Technologies the Biosearch Technologies oKtopure. Validated purifications higher throughput than comparable purification automated Oktopure platform. sbeadex purification protocols have been developed technologies. • Automation-friendly: suitable for most popular for the following plant species (Table 3): Kleargene The advantages of Kleargene include: magnetic-based robotic platforms Plant species with validated sbeadex • Tailor-made lysis conditions according to purification protocols customer requirements Kleargene purification chemistry uses spin- • High throughput DNA purifications (96-well • Quicker results: purification-time optimisation Aphid, soybean Cucumber Rice, Asian (Oryza plates and a glass-fibre support to purify DNA and 384-well formats) • Flexible batch sizes and kit volumes (Aphis glycines) (Cucumis sativus) sativa) using polar interactions to generate PCR-grade • Semi-automated (via Biosearch Technologies • No organic solvents in final wash buffer: results Aubergine Pepper (Capsicum Sunflower (Solanum annuum) (Helianthus DNA, applicable with high throughput methods. Genespin platform) in higher purity melongena) annuus) Protocols have been optimised for both plant and • Manual high throughput purifications (via • No salts in eluates: high Barley Tomato OD260/230 (Hordeum Potato (Solanum (Solanum animal purifications, and the Kleargene chemistry centrifugation) vulgare) tuberosum) lycopersicum) can generate yields between 100 ng and 10 µg • Broad range of sample types (plant leaves, sbeadex feature Specification Rapeseed/Canola/ DNA from samples ranging from plant leaf tissue plant seeds, animal tissue) Colour Dark brown Beet, sugar Oilseed (Brassica Wheat (Triticale) and plant seeds to rodent tails (Figure 4). • High yield of DNA (PCR-grade) (Beta vulgaris) napus) Superparamagnetic Proprietary double-coated bead structure technology (competitor: single- Table 3: Examples of plant species, which have validated purification Amount of coated) protocols using sbeadex chemistry. Kit Application areas starting material Expected yield Irregular Size and shape 80% 5-10 µm (diameter); a; <53 µm Amount Kleargene spin Faster discovery of new traits resulting >6 leaf punches 5 µg Kit Application of starting Expected Table 1: Scales 96-well plates in faster to market. No laboratory at your Maximum capacity 2 µg DNA/mg sbeadex particle areas material yield of available Kleargene spin location required 2-3 leaf punches 1.5 µg Binding material and 2-step binding mechanism: first Kleargene kits. 384-well plates technology silica and second DNA adaptor sbeadex mini 20 – 30 mg 2 µg Purification format 96 plant Plant leaves; generation sequencing applications can be met sbeadex Plant seeds Next-generation ® Automation KingFisher (lab validated); applicable maxi 80 – 100 mg 10 µg by Biosearch Technologies portfolio of sbeadex with alternative open liquid handlers plant sequencing-grade chemistries. All sbeadex kits use surface modified Blood; plant; livestock; forensics; superparamagnetic particles that bind nucleic Sample material plasmids; other human samples Table 4: Scales of available sbeadex plant kits. acids via a novel, two-step binding mechanism ® ® purifications PCR; KASP ; BHQ Probe (Figure 3). Firstly, the nucleic acids affix to the Application chemistry; Sanger; NGS; beads via polar interactions, and secondly, the Microarrays sbeadex nucleic acids are bound via an affinity-driven Storability >12 months mediator. This second binding step allows the final Table 2: Features and specification of sbeadex beads. The requirement to generate high quality and wash steps to be carried out using water-based quantity DNA for PCR, sequencing and next- wash solutions, eliminating the sbeadex chemistry can be used for a variety of species, by modifying the lysis conditions. Although Figure 3: Standard work our standard protocols already deliver high quality flow for all sbeadex-based DNA, sbeadex can be customised and optimised purification chemistry according to customer requirements. 4 5 sbeadex livestock DNA/RNA/total The majority of NGS-based genomic analysis technologies (including those working with livestock samples) typically require high-quality nucleic acid purification DNA preparations which are often challenging and labour-intensive to produce. sbeadex livestock is based on one protocol which can chemistries be used for all sample types and the oKtopure can be used for all livestock samples (excluding blood). MasterPure • Genomic DNA and cDNA cloning • PCR, qPCR, RT-PCR and qRT-PCR • Microarray analysis (including CGH and gene sbeadex blood Some applications require flexibility of isolating expression profiling) Sample type Animal species high-molecular weight genomic DNA, total Bovine Blood samples can often vary considerably in cellular RNA or total nucleic acid from the same The kit uses a scalable salt-precipitation protocol Blood* Chicken sample. The MasterPure™ Complete DNA and which eliminates the need for hazardous Hair follicle Dog quality, and this can lead to wide variations in the RNA Purification Kit allows for isolation of NGS- chemicals (no need for phenol, chloroform or Saliva Fish consistency and yield of extracted DNA. sbeadex Semen Goat blood provides high yields of high-molecular grade DNA, RNA or total nucleic acid in 30–60 guanidine). Depending on the amount of nucleic Tissue (e.g. ear punch) Horse minutes using a single kit, and is suitable for many acid required, and size of the sample, different Buccal swab Sheep weight, double-stranded DNA from different different sample types. MasterPure kits are available, including: FTA cards Pig blood preparations including those treated with anticoagulants, including: Applications for the MasterPure kit include: • MasterPure Complete DNA and RNA Table 5: Sample types from different species applicable with the sbeadex • Library preparation for next-generation Purification Kit livestock kit. *Not compatible with the Oktopure. • EDTA blood • Heparin blood sequencing of genomic DNA and RNA • MasterPure DNA Purification Kit for Blood • Citrate blood • DNA methylation studies (for example, with use • MasterPure Gram Positive DNA Purification Kit ® ® • Buffy coat preparations with the Illumina Infinium HumanMethylation • MasterPure Yeast RNA Purification Kit BeadChips) Application Amount Expected Kit areas of starting yield Large volume DNA purifications material sbeadex Whole blood; Biobanking and other human research markets offers two cost-effective, easy and rapid methods for blood buffy coat 100-200 µL 4-12 µg require large volume purification technologies which the purification of DNA from large volume samples preparations provide high quality DNA. Biosearch Technologies (Table 9): Table 6: Scales of available sbeadex blood kits. Features Kleargene XL PLUS XL Detergent-driven cell lysis, followed by DNA cell-components are enriched, and guanidine-based purification with DNA residual proteins are digested away. Technology binding to silica particles. Contaminants are The DNA is subsequently precipitated, removed using organic solvent-based wash washed, dried, and then resuspended. steps, followed by elution. Automation Manual purifications only Manual purifications only Applications NGS-grade NGS-grade Starting volume Scalable: Up to 10 mL Scalable: Up to 10 mL Sample types Whole blood; Saliva (including Oragene Whole blood; Saliva (including Oragene collection tubes); Tissue; collection tubes); Tissue; Table 7: Key features Typical DNA yields 10-800 µg 10-800 µg of large volume DNA purification chemistries. 6 7
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