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PCR Buffer Set Buffer Set for Optimization of the Magnesium Concentration for the Polymerase Chain Reaction (PCR)

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PCR Buffer Set Buffer Set for Optimization of the Magnesium Concentration for the Polymerase Chain Reaction (PCR) For life science research only. Not for use in diagnostic procedures. PCR Buffer Set Buffer set for optimization of the magnesium concentration for the polymerase chain reaction (PCR) Cat. No. 11 699 121 001 y Version 07 2 × 1 ml each Content version: September 2004 Store at Ϫ15 to Ϫ25° C Product overview Ordering Information Contents Vial Label Content Cat. No.of Roche Applied Science offers a large selection of single enzymes, reagents, and systems for PCR and RT-PCR reagent assays. For a complete overview of our products and ϫ for more detailed information on PCR and RT-PCR 1 MgCl2 •2 1 ml 11 699 113 001 please visit and bookmark our Amplification Special Stock • 25 mM MgCl2 Interest Site at http://www.roche-applied-sci- Solution ence.com/PCR. 2 PCR Buf- •2 ϫ1 ml 11 699 105 001 fer without • Buffer composi- Mg2+, tion (10 ϫ PCR Product Selection Guide 10 × conc. conc.): 100 mM can be viewed at http://www.roche-applied-sci- Tris-HCl, 500 ence.com/sis/amplification/optimal_products/ mM KCl, pH 8.3 optimal_pcr.htm (20°C) Product Pack size Cat. Nos. Application The PCR buffer without MgCl in combination with the Water, PCR Grade 25 ml 03 315 932 001 2 (25 vials of 1 ml) MgCl2 stock solution is used for the individual adjust- ment of the Mg2+ concentration in the PCR reaction. 25 ml 03 315 959 001 (1 vial of 25 ml) In most applications a concentration of 1.5 mM MgCl2 100 ml 03 315 843 001 will yield satisfactory results with a dNTP concentration (4 vials of 25 ml) of 200 μM each. High Pure PCR Template 100 purifications 11 796 828 001 In many cases, however, it s required to titrate the opti- Preparation Kit 2+ mal Mg concentration to increase the specificity and High Pure PCR Product 50 purifications 11 732 668 001 the yield of the PCR reaction. Purification Kit 250 purifications 11 732 676 001 Quality Control Each lot of buffer is function-tested in PCR. PCR Nucleotide Selection No exo- and endonuclease activity is detectable. Storage/Stability The unopened reagents are stable at Ϫ15 to Ϫ25°C Product Description Pack size Cat. No. until the control date printed on the label. Set of Deoxy- Separate vials of dATP, 4 ϫ 25 μmol 11 969 064 001 2+ Nucleotides, PCR dCTP, dGTP, and dTTP. (250 μl) Mg2+ titration Mg ions form soluble complexes with dNTP and template DNA to produce the actual substrate that the Grade 100 mM each polymerase recognizes. PCR Nucleotide Premixed ready-to-use 200 μl 11 581 295 001 The concentration of free Mg2+ ions depends on the Mix solution of PCR Grade 10 ϫ200 μl 11 814 362 001 concentration of compounds like dNTP, free pyrophos- dATP, dCTP, dGTP, and phates (PPi) and EDTA (e.g. from TE buffer). The opti- dTTP. 10 mM. 2+ mal Mg concentration should be determined PCR Nucleotide Premixed ready-to-use 2 ϫ100 μl 11 888 412 001 empirically. It may vary from 1 mM to 5 mM. MixPLUS solution of PCR Grade 11 814 362 001 The most commonly used MgCl2 concentration is 1.5 dATP, dGTP, dTTP mM with a dNTP concentration of 200 μM each. Excess (each 10 mM), and 2+ Mg in the reaction can increase nonspecific primer dUTP (30 mM) binding and increase the non-specific background of the reaction. 2+ Trademarks All product names and trademarks are the property of Too little Mg in the reaction can result in a lower their respective owners. yield of the desired product. The following table is a guideline for adjusting the MgCl2 concnetration. In a 50 μl PCR reaction use 5 μl PCR reaction buffer without MgCl2 and the respective amount of MgCl2:. MgCl2 concentration 1 mM 1.25 mM 1.5 mM 1.75 mM 2 mM 2.5 mM 5 mM Contact and Support Volume of MgCl2 2 μl 2.5 μl 3 μl 3.5 μl 4 μl 5 μl 10 μl stock solution To ask questions, solve problems, suggest enhancements or report new applications, please visit our Online Technical Support Site at: www.roche-applied-science.com/support Changes to • Regulatory Disclaimer updated To call, write, fax, or email us, visit the Roche Applied Science home page, previous version www.roche-applied-science.com, and select your home country. Country- specific contact information will be displayed. Use the Product Search func- tion to find Instructions for Use and Material Safety Data Sheets. Roche Diagnostics GmbH Roche Applied Science 68298 Mannheim Germany 0412.11714 180001➆.
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