ACCCACGAAAGGGAA ATAAGC

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1 Lyophilisation Endpoint PCR Real-Time PCR Reverse Transcription dntps Custom-Specific Lyophilisation ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCAC- GAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO Molecular Biology

2 2 Introduction Building Blocks of Life No fridge required. ready-to-use individual scales customized packaging & labeling

3 Introduction 3 Building Blocks of Life Nucleotides & Nucleosides In our chemistry division, we have hundreds of natural and modified nucleotides in stock. In addition, with our pre-made building blocks and in-house expertise we mavnufacture even the most exotic nucleotide analog from mg to kg scale. Click Chemistry, Probes & Epigenetics Our Probes & Epigenetics as well as Click Chemistry sections offer innovative reagents for the functionalization, conjugation and labeling (fluorophores, haptens) of (bio) molecules complemented by epigenetic modification analysis tools. LEXSY Expression In the field of recombinant protein production, Jena Bioscience has developed its proprietary LEXSY (Leishmania Expression System) technology. It is based on an S1-classified unicellular organism that combines easy handling with a eukaryotic protein folding and modification machinery. Besides everything you need to establish LEXSY in your lab we also offer custom expression of recombinant proteins. Crystallography & Cryo-EM For the crystallization of biological macro-molecules which is often the bottleneck in determining the 3D-structure of proteins we offer specialized reagents for protein stabilization, crystal screening, crystal optimization, and phasing that can reduce the time necessary to obtain a high resolution protein structure from several years to a few days. Molecular Biology & Proteins For applications in the field of Molecular Biology we offer a large selection of single reagents, complete kits and optimized master mixes. This section includes products for DNA or RNA purification, amplification and modification with focus on PCR-related techniques. For your questions regarding lyophilisation contact me directly: pcr@jenabioscience.com

4 4 Building Blocks of Life Established in 1998 by a team of scientists from the Max-Planck-Institute of Molecular Physiology (Dortmund), Jena Bioscience utilizes more than 25 years of academic know-how to develop innovative reagents for clients from both research and industry in 100+ countries. To date, Jena Bioscience still remains an owner-operated business. Imprint: Design & Layout by timespin - Digital Communication GmbH, Sophienstr. 1, D Jena, Germany, Copyright: Please contact Jena Bioscience if you want to use texts and/or images in any format or media. For the sake of the environment: this brochure is printed on chlorine-free natural paper.

5 ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA Introduction ATAAGC AACO 5 TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO Contents Introduction...6 Endpoint PCR... 7 Real-Time PCR...8 Reverse Transcription...9 dntps...10 Custom-Specific Lyophilisates... 11

6 6 Introduction Building Blocks of Life Introduction Good to know Figure 1 Phase Diagram of Water Lyophilisation (= freeze-drying) is a dehydration method applying low temperatures and reduced ambient pressure. In these conditions water sublimes from a solid state directly into the gas phase (Fig. 1). As a result, the moisture content of a product is decreased to approximately 1 % [1]. In the absence of water growth of microorganisms and chemical (degradation) reactions are inhibited leading to long-term preservation. Freeze-dried materials may be stored at ambient temperatures for many years while remaining structurally & chemically stable. They are fully reconstituted after addition of water (rehydration). When to consider freeze-drying? Modern freeze-drying techniques revolutionized preservation of perishable materials not only for chemical or food industries. Lyophilisation also provides significant advantages to many of todays molecular biological applications and products: 1. Due to the mild operating conditions (low pressure & temperatures; Fig. 1) freeze drying is particularly suitable for (heat-) sensitive products (e.g. enzymes) 2. Lyophilisation dramatically enhances shelf life, which is especially useful for products with poor stability in aqueous solutions (e.g. proteins) 3. Since dry mixes require no refrigeration, they are stored and handled at room temperature. At the same time, no activity loss caused by freeze-thaw cycles occurs. Lyophilisate Liquid Mix Figure 2 Jena Bioscience in-house data show that freeze-dried PCR mixes remain fully functionals after 12 month heat exposure (30 C, green circle) while liquid mixes significantly lose activity (4 C, red circle). (Assay: Amplification of a 4 kb fragment, lambda DNA, template dilution series) Reference: [1] Patel et al (2011) Emerging freeze-drying process development and scale-up issues. Aaps Pharmscitech 12 (1):

7 Endpoint PCR 7 Endpoint PCR Preloaded and stable at Room Temperature Jena Bioscience lyophilisates are ready-to-use and preloaded in a dry, room temperature stable format. Mixes are available in preferred packaging such as: single PCR reaction tubes 8-tube strips (low/high profile) 96-well plates (low/high profile; flat top, half/full skirt) custom-specific formats Direct PCR Lyophilisates are designed for amplification of target DNA directly from whole blood, animal and plant tissues. No pretreatment or prior purification of DNA is required. PCR-160S 192 reactions x 20 µl 200,00 PCR-160L 960 reactions x 20 µl 800,00 Multiplex PCR Master Lyophilisate is especially designed for parallel amplification of a variety of fragments in a single PCR assay. The mix is suitable for multiple target gene amplification in a single tube. PCR-161S 192 reactions x 20 µl 212,50 PCR-161L 960 reactions x 20 µl 850,00 Crystal Taq Master Lyophilisate recommended for routine PCR applications (up to 4 kb fragment length), high throughput PCR or genotyping. PCR-152S 192 reactions x 20 μl 93,90 PCR-152L 960 reactions x 20 μl 375,60 Crystal Hot Start Master Lyophilisate recommended when amplifying low-copy-number targets, working with complex backgrounds or when prolonged room-temperature set-ups are unavoidable. PCR-153S 192 reactions x 20 μl 114,70 PCR-153L 960 reactions x 20 μl 458,80

8 8 Real-Time PCR Building Blocks of Life Real-Time PCR No freeze-thaw cycles No more pipetting on ice. Jena Bioscience Real-Time PCR lyophilisates are designed for quantitative DNA analysis in real-time based on flourescent DNA stains or labeled probes. All mixes are also available in combination with high ROX as reference dye for cycler-internal signal normalisation. JBS qpcr ProbesMaster Lyophilisate is based on detection of labeled DNA probes. Mixes are recommended for use with Dual Labeled Fluorescent Probes, e.g. TaqMan, Molecular Beacons or FRET probes. qpcr ProbesMaster Lyophilisate PCR-156S 192 reactions x 20 μl 137,50 PCR-156L 960 reactions x 20 μl 550,00 qpcr ProbesMaster Lyophilisate highrox PCR-168S 192 reactions x 20 µl 137,50 PCR-168L 960 reactions x 20 µl 550,00 qpcr GreenMaster Lyophilisate is based on fluorescent DNA stain fluorescent DNA stain EvaGreen. No sequence-specific labeled probes are needed. qpcr GreenMaster Lyophilisate PCR-157S 192 reactions x 20 μl 137,50 PCR-157L 960 reactions x 20 μl 550,00 qpcr GreenMaster Lyophilisate highrox PCR-170S 192 reactions x 20 µl 137,50 PCR-170L 960 reactions x 20 µl 550,00 qpcr SybrMaster Lyophilisate is based on fluorescent DNA fluorescent DNA stain SYBR Green. No sequence-specific labeled probes are needed. qpcr GreenMaster Lyophilisate PCR-173S 192 reactions x 20 µl 137,50 PCR-173L 960 reactions x 20 µl 550,00 qpcr SybrMaster Lyophilisate highrox PCR-174S 192 reactions x 20 µl 137,50 PCR-174L 960 reactions x 20 µl 550,00

9 ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC Reverse ACCCACGAAAGGGAA Transcription ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO 9 Reverse Transcription One-Step RT-PCR Master Lyophilisate is designed for performing maximum sensitive and specific RT-PCRs convenient in single tubes. The enzyme mix is based on a genetically engineered RT polymerase with enhanced thermal stability resulting in an increased specificity, higher cdna yield and an improved efficiency for highly structured and long cdna fragments. PCR-159S 192 reactions x 20 μl 475,00 PCR-159L 960 reactions x 20 μl 1.900,00 RT Master Lyophilisate contains a genetically engineered version of M-MLV Reverse Transcriptase (M-MLV RT) with eliminated RNase H activity and increased thermal stability. The enzyme is a RNAdirected DNA polymerase that synthesizes a complementary DNA strand initiating from a primer using single-stranded RNA or DNA as template. PCR-158S 192 reactions x 20 μl 370,00 PCR-158L 960 reactions x 20 μl 1.480,00

10 10 dntps Building Blocks of Life dntps Jena Bioscience's enzymatic dntp manufacturing process ensures superior quality for deoxynucleotide triphosphates: All our sodium salt dntp lyophilisates are ultrapure (>95% by HPLC) and functionally tested by a set very sensitive enzymatic reactions. datp - Solid dctp - Solid dgtp - Solid dttp - Solid dutp - Solid ditp - Solid NU μmol 20,46 NU x 20 μmol 81,84 NU μmol 20,46 NU x 20 μmol 81,84 NU μmol 20,46 NU x 20 μmol 81,84 NU μmol 20,46 NU x 20 μmol 81,84 NU μmol 24,55 NU x 20 μmol 98,21 NU μmol 24,55 NU x 20 μmol 98,21 Equipment There is a great diversity of freeze-dryers on the market. At the most basic level however, they all share 4 essential components: Chamber (to hold samples) Refrigerating system (to cool down samples) Vacuum pump (to reduce chamber pressure) Condenser (to remove sublimed water) Jena Biosciences uses a compact two-chamber freeze-dryer from Zirbus Technology (Figure 3). The microprocessor-controlled system enables flexible adjustment of desired process parameters (e.g. shelf temperature, chamber pressure) and continuous documentation via its internal Zirbus Dry Control operating and recording tool. Figure 3 Foto by courtesy of Christian Zirbus, Zirbus technology GmbH

11 Custom-Specific Lyophilisates 11 Custom-Specific Lyophilisates Looking for freeze-dried mixes beyond the standard Jena Bioscience repertoire? But you are reluctant to take the high costs of equipment acquisition? you need a reliable partner with the necessary technical advice and know how? We manufacture individualized lyophilisates according to your requirements (including addition of primers or DNA controls) and applications (Endpoint PCR, Real-Time PCR, Reverse Transcription). Liquid Mix Based on your liquid mixes we develop formulations, lyophillise at desired scales and offer customized product packaging & labeling. 1. Formulation Development 2. Small-scale testing - Functionality - Stability - Appearance Pilot-Scale Optimization Scale-Up Quality Control Custom - Dispensing 3. Re-formulation - Packaging - Labeling - Stabilizers - Protectors - Process parameters Individual freeze-dried mix Manufactured under ISO 9001 reagulations and validated by customer on-site audits Jena Bioscience guarantees long-term product stability ( 6 months at 30 C). me for service inquiries or further information: pcr@jenabioscience.com

12 Jena Bioscience GmbH Loebstedter Str Jena Germany Phone +49 (0) Fax +49 (0) Test the exceptional performance of our lyophilisates for yourself. Request a sample! Our entire product range is available as samples. ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCAC- GAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO Molecular Biology

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