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Molecular Biology - Nucleic Acids PCR in end point Hot Start DNA polymerases

Hot Start DNA polymerases

Biodyne HOT FIREPol®

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tiAmplus DNA Polymerase

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• Optimized mixture of thermostable DNA polymerases Taq (Thermus aquaticus), Pfu (Pyrococcus furiosus) and a polymerization activator
• Ultrapure recombinant polymerases, thermostable and hot start. Pfu proofreading
• The hot start property of the polymerases makes them inactive at room temperature allowing the preparation of the reaction mix at room temperature
• Unique composition of the reaction buffer promoting pH stability at high temperature and ensuring high efficiency for long sequence amplifications
• Amplification of genomic DNA sequences from 3 to 25 kb and episomal DNA sequences up to 40 kb
• Particularly suitable for the amplification of eukaryotic genomes
• Ideal for genome mapping and sequencing of long fragments
• Supplied with a 10X reaction buffer
• Available in 100 and 500 unit formats

Cytiva Hot Start MasterMix

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• For demanding PCR applications in which high specificity are essential
• Reduces nonspecific priming
• Kit Contents: Tris-HCl, KCl, MgCl2, dNTPs, Taq DNA Polymerase, Hot Start Activator protein, additional MgCl2 tube

Cytiva PCR illustra™ kits: Ready-To-Go™ technique

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The RTG (Ready-To-Go) is to provide the mix necessary for the reaction as preformulated, premixed, predispensed beads

• Increased reproducibility
• Minimized pipetting steps
• Reduced the potential for contamination
• Long-term ambient-temperature stable (> 12 months)

Illustra Hot Start Mix RTG beads: for PCR requiring high amplification specificity. The only reagents to be added are water, the DNA to amplify and the primers.

Polymerase Chain Reaction (PCR) is covered by patents held by Roche Molecular Systems and F Hoffmann-La Roche Ltd.

SolisFAST ADN Polymerase

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• For amplification in fast PCR mode, reduces the time of a PCR cycle by up to 4 times compared to a standard protocol.
• Elongation step 3 to 4 times faster than with conventional DNA polymerase
• Hot start enzyme inactivated at room temperature, activation in only 30 seconds at 98 °C
• For the amplification of Low complexity DNA fragments (cDNA, lambda, plasmid DNA) up to 5 kb or up to 3 kb for human, animal or plant DNA
• Ready-to-use mix Kit; requires only the matrix, primers and molecular biology quality water
• Available in a 'Ready to load' version for direct deposit on gel, contains a load buffer and two dyes for migration monitoring (blue (3,5-4,5 kb) and yellow (35-45 pb))
• Available in a version with UNG to reduce the risk of cross-contamination
• Stable 30 days at room temperature, long-term storage at -20 °C

qPCR SolisFAST Mix

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• Mix for quantitative PCR with fast protocol, elongation 3 to 4 times faster than with conventional DNA polymerase
• Hot start enzyme inactivated at room temperature, activation in only 30 seconds at 95 °C
• qPCR results obtained in only 30 minutes depending on thermocycler performance (optimized for qPCR Mic thermocyclers (Cat No. 634001 and 634002)
• Mix also available with ROX or PURPLE dye for thermocyclers requiring a reference signal for data normalization.
• Stable 30 days at room temperature, long-term storage at -20 °C

OnTaq DNA Polymerase Hot Start Kit (2.5 U/µl) with dNTPs

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• DNA polymerase hot start at an initial concentration of 2.5 U/µl
• Increases PCR specificity, sensitivity and throughput by reducing the risk of mismatch and primer-dimer formation
• Activated during the initial 10 min denaturation step at 95 °C
• Conserved 5'-'3' exonuclease activity and absence of 3'-'5' exonuclease activity
• Adds a free adenine to the 3' end of the amplicon (monoadenylation)
• Kit with 3 different 10X reaction buffers: without MgCl2 (A), with MgCl2 (B), with 2 inert dyes for direct deposit (C)
• Kit supplied with dNTPs (separate tube)

Fastgene ApTaq Hot Start DNA polymerase

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• DNA polymerase Taq hot start
• Hot start system by aptamer technology: reversible activation / inactivation
• Polymerase inactivated below 45 °C
• Fully activated polymerase above 60 °C
• No specific activation step required as with antibody technology
• Ideal for rapid PCR, routine PCR and SNP analysis in particular

Master mix Fast Sybr Green qPCR (2X) and Master mix Fast SG qPCR (2X) with ROX solution

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• Fast qPCR mix with SYBR Green I dye for use with most real-time thermal cyclers
• Contains Perpetual Taq hot start DNA polymerase, optimised buffer, dNTPs (dTTP partially replaced by dUTP) and SYBR Green I dye
• Uracile N-glycosylase (UNG) heat-labile provided to limit the risk of cross-contamination (optional use)
• Available with ROX dye (separate tube) for standardisation

Master mix Fast Probe qPCR (2X) and Master mix Fast Probe qPCR (2X) with ROX solution

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• Fast qPCR mix with hydrolysis probes for use with most real-time thermal cyclers
• Contains Perpetual Taq hot start DNA polymerase, optimised buffer, dNTPs (dTTP partially replaced by dUTP) and SYBR Green I dye
• Uracile N-glycosylase (UNG) heat-labile provided to limit the risk of cross-contamination (optional use)
• Available with ROX dye (separate tube) for standardisation

Master mix HRM PCR (2X)

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• Mix designed for high resolution melting curve (HRM) analysis of DNA samples
• Allows detection of gene mutations and single polymorphisms (SNPs) including class III and IV
• Contains onTaq hot start DNA polymerase, optimised buffer, dNTPs and fluorescent dye
• OnTaq polymerase activated during the initial denaturation step of the PCR at 95 °C for 15 min
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