Molecular biology , Proteins and Reagents

Custom Services and Solutions www.biotechrabbit.com Headquarters Subsidiary Commercial Register: Neuruppin biotechrabbit GmbH biotechrabbit GmbH (HRB 9381 NP) Neuendorfstr. 24a Volmerstr. 9 Managing Director: Dr. Bernd Haase 16761 Hennigsdorf, Germany 12489 Berlin, Germany Principal Office: Hennigsdorf [email protected] [email protected] Tax Office: Oranienburg www.biotechrabbit.com www.biotechrabbit.com VAT no/USt-Id: DE 279969868 Office: +49 3302 20754 10 Office: +49 30 555 7821 10 Tax no: 05310604935 Fax: +49 3302 20754 11 Fax: +49 30 555 7821 99

2 Table of Contents

About biotechrabbit 4

CAPITAL qPCR™ Probe Mix 6

CAPITAL qRT-PCR™ Probe Mix 6

CAPITAL qRT-PCR™ Green Mix 7

One Step RT-PCR Mix 7

Direct-Load Hot Start PCR Mix 8

ApStarTaq™ DNA Polymerase 8

UPstart™ Anti-Taq Antibody 9

Heat-Labile Uracil–DNA Glycosylase 9

biotechrabbit product portfolio 10

3 About biotechrabbit

biotechrabbit innovates, develops and manufactu- res products with superior per- formance for diagnostics, life science research and applied markets. All development, production and logistics take place in our Berlin facility.

Molecular biology and protein production is the cornerstone of biotechrabbit’s offering. We use high density prokaryotic and eukaryotic fermentati- on followed by several fine-tuned purification steps to deliver highly pure proteins. Capacities range from small to large-scale production.

The research and development department with experienced scientists and business developers is emphasizing on customers and partners‘ needs. The team is highly engaged in providing top-quality pro- ducts for molecular biology applications.

4 ISO 13485:2016 and ISO 9001:2015 certification ex- emplify biotechrabbit’s commitment to the highest quality standards. The scope of our quality manage- ment includes: design and development, production, marketing and distribution of molecular biology pro- ducts, proteins and reagents for in-vitro diagnostics.

Certified Quality-System

Quality control analyses after each key production step secure product performance. Sophisticated ac- tivity analysis in conjunction with purity, concentrati- on and contamination tests confirm the consistently high quality of all biotechrabbit products.

Lyophilization as a potential final processing step fulfills the continually growing demand for product stability without depending on a temperature-con- trolled supply chain. Developed for low hygrosco- picity combined with high rigidity, biotechrabbit lyo cakes and beads perfectly match all stability and performance requirements.

5 CAPITAL qPCR™ Probe Mix

Best in-class performance for both single- and multiplex targets Convenient master mix with high specificity in pathogen detection Highly sensitive for low-abundance DNA targets biotechrabbit CAPITAL qPCR Probe Mix for quantifying genomic, cDNA and viral sequences provides outstanding performance in single and multiplex qPCR. The high sensitivity provided by the mix is ideal for detection of low-abundance DNA targets in applica- tions, such as pathogen detection.

A 6000 7000 36 34 Pathogen no. 1 2 3 4 32 5000 Ct BTR 21.75 20.31 22.86 21.90 6000 30

Cq 28 Ct vendor T N/A 26.21 37.34 40.64 BTR 26 4000 5000 24 E = 99.8% R2=0.990 Slope=-3.327 y-int=41.072 22 2 3 4 5 6 3000 4000 Log Starting Quantity RFU RFU 3000 hgDNA 200 ng 20 ng 2 ng 0.2 ng 0.02 ng 2000 Ct BTR 22.59 26.15 28.81 32.61 36.48 T 2000 Ct vendor T 24.09 26.94 30.25 33.73 37.71 1000

1000 0 0 0 10 20 30 40

0 10 20 30 40 Cycles Cycles BTR Vendor T B 2500 Accurate quantification over a wide linear range. A qPCR assay was Pathogen no. 1 2 3 4 Ct BTR 34.23 32.15 35.73 35.59 BTR performed to evaluate the sensitivity, specificity and absence of non- 2000 Ct vendor T 44.54 37.34 N/A N/A specific amplification in non-template controls. Human genomic DNA was 1500 diluted (200 ng to 0.02 ng) and amplified using the CAPITAL qPCR Probe RFU Mix and a mix from Vendor T. 1000

High sensitivity for pathogen detection in multiplex assays. A 4-plex 500 qPCR assay for detecting human pathogens was performed using T ­CAPITAL qPCR Probe Mix and a mix from Vendor T. The biotechrabbit 0 mix demonstrated reliable and sensitive detection with both high- and 0 10 20 30 40 low-abundance templates (panels A and B, respectively). Cycles CAPITAL qRT-PCR™ Probe Mix

Best in-class performance for single and multiplex reactions Convenient master mix for detection of low-copy pathogen targets High specificity and sensitivity across a wide range of sample sources biotechrabbit CAPITAL qRT-PCR Probe Mix provides outstanding performance for real-time PCR quantification of RNA templates, including mRNA, total RNA and viral RNA from a wide range of targets. The proprietary master mix ensures high specificity and sensitivity with one or more targets, making it the choice for extremely low-copy-number targets in pathogen detection.

3000 12

High Specificity: No signal with 2500 non-template control (including 10 500 ng Human gDNA) RFU 2000 8 ) 3

102 1500 6 0 10 20 30 40 Cycles RFU RFU (10 1000 4

500 2

0 0

0 10 20 30 40 0 10 20 30 40 Cycles Cycles BTR Vendor N

Superb multiplexing performance: high sensitivity and specificity. CAPI- Sensitive and robust RT-qPCR. A one-step RT-qPCR assay was perfor- TAL qRT-PCR Probe Mix was used to perform 3-plex, one-step qPCR med to evaluate low-input detection, specificity and absence of non- for detecting duplications of three different viral RNA targets. The mix specific amplification in non-template control. RT-qPCR targets were performed with high specificity and excellent accuracy in multiplex de- quantitated with 10 ng to 1 pg total RNA targeting GAPDH. biotechrabbit tection. CAPITAL qRT-PCR Probe mix showed earlier Ct in all dilutions compared 6 to vendor N. CAPITAL qRT-PCR™ Green Mix

Convenient mix for quantification of RNA templates Sensitive and specific amplification Excellent linearity across a wide range of RNA dilution biotechrabbit CAPITAL qRT-PCR Green Mix allows sensitive and specific cDNA synthesis and qPCR in a single tube for quantifying mRNA, total RNA and viral RNA sequences. Extremely low-copy-number targets can be detected with high efficiency over several logs of template concentration. The proprietary CAPITAL qRT-PCR Green technology is provided as a convenient master mix.

30 BTR Melt Peak Vendor P

3000 Vendor N 25

2000

20 -d(RFU)/dT 1000 ) 3 0 15 60 70 80 90 Temperature, Celsius RFU (10 P NTC 10

5

BTR NTC 0

0 10 20 30 40 Cycles

Superb sensitivity and specificity in RNA quantification. One step RT-qPCR was performed with a serial dilution of human total RNA (10 ng to 1 pg per reaction) to evaluate the CAPITAL qRT-PCR Green Mix and commercially available dye-based QRT-PCR reagents. The biotechrabbit mix exhibited robust amplification with high specificity, earlier Ct and higher signal than the mixes from two other vendors.

One Step RT-PCR Mix

Fast and convenient endpoint PCR assay setup High yield of amplification Highly specific detection biotechrabbit One Step RT-PCR Mix provides fast and convenient setup for reverse transcription and amplification in a single step. The blend of efficient thermostable reverse transcriptase and proprietary Ribonuclease Inhibitor ensures high cDNA yields. The optimized composition allows highly specific detection, even of complex templates.

High-yield amplification. cDNA was synthesized from 500 ng human total RNA in a 50 µl reaction at 50˚C and subjected to 40 cycles of amplification of a 111 bp product using the One Step RT-PCR Mix and a kit from Vendor P. The biotechrabbit mix pro- duced high yields and exhibited no amplification of side products, illustrating its high specificity. NTC: no template control. 7 Direct-Load Hot Start PCR Mix

Room-temperature reaction setup and direct gel loading High specificity and sensitivity Amplification of low abundance targets with high yield biotechrabbit Direct-Load Hot Start PCR Mix contains tracking dyes and allow reactions to be loaded directly into agarose gels without the addition of loading buffer. The hot-start mix provides room-temperature reaction setup and highly specific and sensitive amplification for high yields even with low-abundance targets.

A B

25 ng template 2.5 ng template

High-yield amplification of a low-abundance target. A 750 bp target was amplified from dilutions of human genomic DNA (per reaction: 25 ng, Panel A; 2.5 ng, Panel B) and loaded directly onto an agarose gel along with a 1 kb ladder (M). The biotechrabbit mix produced higher yields, especially at low template concentrations, compared to mixes from Vendors P and K. ApStarTaq™ DNA Polymerase

Instant enzyme activation High sensitivity, specificity and yield in real-time and endpoint PCR Convenient for room temperature assay setup biotechrabbit ApStarTaq DNA Polymerase is an -based hot-start enzyme that ensures high amplification yields for real-time and endpoint PCR. Convenient room-temperature setup is provided by an aptamer which inhibits Taq DNA Polymerase at ambient temperatures. Activity is instantly restored at 45°C for highly sensitive and specific amplification.

7000 hgDNA 200 ng 20 ng 2 ng 0.2 ng 6000 Ct BTR 22.11 25.21 28.08 31.82 Ct vendor R 23.02 26.63 29.85 32.59 5000

4000

RFU 3000

2000

1000

0

0 10 20 30 40 Cycles

BTR Vendor R

Superior results with high-sensitivity qPCR amplification. Human geno- Maximum application convenience with high yield and specificity. A 750 mic DNA was diluted (200 ng to 0.2 ng per reaction) and amplified using bp target was amplified from 2.5 ng human genomic DNA to assess the ApStarTaq DNA Polymerase and an aptamer from Ven- yield and specificity using ApStarTaq DNA Polymerase and a polymerase dor R to evaluate sensitivity and specificity. Results demonstrate distinct- from Vendor R. Higher yield was obtained using the biotechrabbit poly- ly earlier Ct values and higher signal of detection for the biotechrabbit merase. NTC: no template control. polymerase. 8 UPstart™ Anti-Taq Antibody

Efficient Taq DNA Polymerase inhibition and defined release at elevated temperature Increased target yield and specificity with reduced non-specific amplification Exceptionally pure monoclonal antibody produced in cell culture biotechrabbit UPstart Anti-Taq Antibody is an ultra-pure monoclonal antibody providing highly efficient hot-start for PCR, enhancing specificity and sensitivity and improving yields. The antibody binds to Taq DNA polymerase, efficiently preventing amplification before the first denaturation step. At high temperatures, the antibody becomes inactive and releases active Taq DNA polymerase.

Efficient temperature-dependent inhibi- 120,00 tion of Taq DNA Polymerase. Amplifica- tion reactions were pre-incubated with 100,00 Released Taq activity UPstart Anti-Taq Antibody at different temperatures, followed by an isother-

80,00 mal polymerase assay. Taq activity was inhibited up to 70°C and restored after pre-incubation at elevated temperatu- 60,00 res.

40,00 Activity [%] Activity

20,00 Taq inhibition

0,00

-20,00 0 5 10 15 20 25 30 35 40 min Control (Taq) 60°C, 3 min 65°C, 3 min 70°C, 3 min 75°C, 1 min 80°C, 1 min 85°C, 1 min Pre-incubation temperatures

Heat-Labile Uracil–DNA Glycosylase

Extremely beneficial in qPCR and RT-qPCR Inactive at 50°C, entirely inactivated by heating to 80°C Highly purified enzyme free of RNases and DNases biotechrabbit Heat-Labile Uracil–DNA Glycosylase improves qPCR and RT-qPCR performance by selectively degrading carryover contamination with DNA amplification products. The enzyme degrades uracil-containing DNA as initial step before running reverse transcription or PCR amplification. Efficient heat inactivation of this highly pure, RNase- and DNase-free enzyme ensures all target DNA/RNA or PCR product remain intact.

Efficient removal of PCR carryover con-

375,000 tamination. An uracil-containing PCR 350,000 product was used as template (100 ng, 325,000 10 ng and 1 ng per reaction) and subjec- 300,000 ted to qPCR with and without Heat-La- 275,000 bile Uracil–DNA Glycosylase treatment. 250,000 Amplification at all DNA concentrations 225,000 is a result of the simulated carryover 200,000 contamination. The lack of amplification 175,000 in HL-UDG treated reactions indicates ∆Rn 150,000 that ­carryover DNA was successfully 125,000 removed. 100,000

75,000

50,000

25,000

0

-25,000 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 Cycle No HL-UDG treatment HL-UDG pre-treated No template control

9 biotechrabbit product portfolio

Real-time PCR CAPITAL qPCR™ Probe Mix CAPITAL qPCR™ Green Mix CAPITAL qRT-PCR™ Probe Mix CAPITAL qRT-PCR™ Green Mix

DNA Polymerases and Master Mixes Taq / Hot Start Taq DNA Polymerase and PCR Mixes ApStarTaq™ DNA Polymerase Direct-Load Hot Start PCR Mix HiFi DNA Polymerase Long Range Hot Start PCR Mix

Reverse Transcription Reverse Transcriptase One Step RT-PCR Mix cDNA Synthesis Kit

Enzymes and proteins UPstart™ Anti-Taq Antibody T7 RNA Polymerase T4 DNA Ligase Heat-Labile Uracil–DNA Glycosylase Proteinase K RNase Inhibitor

More product lines Lyophilized Polymerases and Mixes Nucleotide Sets and Mixes GenUP™ Nucleic Acid Purification Kits DNA/Protein Electrophoresis Ladders Cell-free Protein Synthesis Kits (RTS) Tris-NTA Amine / Biotin (High affinity His-tag binding)

10 Legal Disclaimer and product use limitation biotechrabbit GmbH makes every effort to include accurate and up-to-date information within this publication; however it is possible that omissions or errors might have occurred. Changes of publication can be made at any time without notice. All mentioned trade- marks are protected by law. The purchase of biotechrabbit product does not include a license to perform any patented applications; therefore it is the sole responsibility of users to determine whether they may be required to engage a license agreement depending upon the particular application in which the products are used. biotechrabbit products are developed, manufactured, and sold for in-vitro use and laboratory use only. They are not suitable for administration to humans or animals. The products are to be used only by trained laboratory personnel who are familiar with potential hazards and in laboratories equipped to perform life science research. biotechrabbit products may contain chemicals which are harmful if misused. Due care should be taken with all products to avoid direct contact with components that are potentially harmful.

Trademarks biotechrabbit™, CAPITAL qPCR™, CAPITAL qRT-PCR™, ApStarTaq™, UPstart™, GenUP™ (biotechrabbit GmbH)

Medica 2018, Düsseldorf, Germany, 12.-15.11.2018 www.biotechrabbit.com © 2018 biotechrabbit GmbH, all rights reserved. PTM-1008-001 valid from 17.10.2018 11 biotechrabbit GmbH, Berlin, Germany [email protected] www.biotechrabbit.com