The highfidelity pcr master mix is in the 2x format. Faststart high fidelity pcr system, dntpack from roche. Efficacy of different dna polymerase enzymes in pcr. System, introduced in 2003, roche applied science created a. Control template and primers are provided for 20 control reactions. Faststart high fidelity pcr system, dntpack faststart high fidelity enzyme blend with additional readytouse 10 mm pcr grade nucleotide mix cat. Pcr combines all the benefits of faststart taq dna polymerase with. The faststart high fidelity pcr system an optimized enzyme blend for hot start pcr combines all the benefits of faststart taq dna polymerase with 4fold higher accuracy and the abilty to amplify fragments up to 5 kb. If cdna synthesis reaction mixture is used as a source of template, the volume of the template should not exceed 10% of the final pcr reaction volume.
Faststart high fidelity pcr system comprises thermostable and. Mar, 2015 fulllength lncrnahit cdna was amplified using the faststart high fidelity pcr system roche applied science and nmumg genomic dna as a template. Phusion highfidelity dna polymerases and master mixes phusion highfidelity dna polymerase 100 u f530s 500 u f530l phusion green highfidelity dna polymerase 100 u f534s 500 u f534l phusion highfidelity pcr master mix with hf buffer 100 x 50 l rxns f531s 500 x 50 l rxns f531l phusion highfidelity pcr master mix with gc buffer. Find safety data sheets, certificates of analysis and method sheets instructions for use in the roche support portal elabdoc in case of questions please contact your local custombiotech representative. Pcr optimization of the control template using phusion highfidelity pcr kit protocol for phusion highfidelity pcr master mix with gc buffer pcr protocol for crimson longamp taq dna polymerase m0326. Faststart high fidelity pcr system catalog number 03553361001. The faststart high fidelity pcr system is the ideal tool for all amplification experiments where a combination of high specificity, sensitivity, accuracy, and yield is needed. Phusion highfidelity dna polymerases and master mixes phusion highfidelity dna polymerase 100 u f530s 500 u f530l phusion green highfidelity dna polymerase 100 u f534s 500 u f. Choose from our high fidelity pcr polymerases whenever accuracy is required, including for challenging reaction conditions such as longrange pcr, fast pcr or amplification of gcrich or. The faststart high fidelity pcr system is a blend of faststart taq dna polymerase and a thermostable proofreading protein, which is also chemically modified. Faststart high fidelity pcr system, dntpack roche elabdoc. Product information thermo scientific highfidelity pcr. Im adding it to the article unless there are any objections.
In a subsequent pcr, 5 l cdna template is used in a total volume of 50 l with the expand long template pcr system to amplify a 10 kb dystrophin fragment. If cdna synthesis reaction mixture is used as a source. Please see the online application note stepbystep protocol for multiplex pcr with the faststart high fidelity pcr system and the pcr optimization kit cat. The faststart high fidelity pcr system is an enzyme blend of chemically modified taq dna polymerase faststart taq dna polymerase and a thermostable via chemical. The system claims to have a 2fold increase in fidelity over the expand high fidelity pcr system and a 6fold increase in fidelity compared to taq dna polymerase alone. Prevent the extension of nonspecifically bound primers using this hot start enzyme. In addition to high accuracy, pfuultra high fidelity dna polymerase provides robust amplification of long, complex genomic targets.
Description the superscriptz iii onestep rtpcr system with platinum. Faststart high fidelity pcr system, dntpack sigmaaldrich. Fortyeight pairs of primers were pooled and diluted to have a. General description the enzyme dntpack comprises faststart taq dna polymerase and a readytouse solution of pcr grade nucleotides. As this note demonstrates, the system amplifies up to 18 pcr fragments simultane. High pure, faststart and expand are trademarks of roche. Superscript iii onestep rtpcr system with platinum taq. Product information thermo scientific highfidelity pcr kit. Highthroughput and sensitive quantification of circulating. For amplification of multiple targets in a single reaction multiplex pcr, faststart high fidelity pcr system offers several advantages. As this note demonstrates, the system amplifies up to 18 pcr fragments simultaneously after the reaction is optimized with the roche applied science pcr optimization kit 2. This enzyme mixture transcribes more accurately up.
The high fidelity pcr master mix is in the 2x format. When the cycle is repeated several times, the net result is a rapid increase in the total number of copies of the target. The faststart high fidelity pcr system is a blend of faststart taq dna. The superscript iii onestep rt pcr system with platinum taq high fidelity is designed for convenient endpoint detection and analysis of rna molecules by onestep rt pcr.
Faststart taq dna polymerase and high fidelity pcr system faststart taq dna polymerase is ideal for single target hot start pcr amplification and multiplex applications. For high complexity genomic dna, the amount of dna template should be 50250 ng per 50. Expandtmlong template 1681 834 100 u pcr system 30 reactions pcr 0. A range of high fidelity, regular, economy and premium taq polymerase enzymes were selected. Easya highfidelity pcr cloning enzyme 5 table i potential pcr cloning errors using taq dna polymerase versus easya highfidelity pcr cloning enzyme pcr product size clones with. Prevent the extension of nonspecifically bound primers. Easya high fidelity pcr cloning enzyme 5 table i potential pcr cloning errors using taq dna polymerase versus easya high fidelity pcr cloning enzyme pcr product size clones with errorsa easya high fidelity pcr cloning enzyme taq dna polymerase 250 bp 0. Dna polymerase in an enzyme blend faststart high fidelity pcr. The roche applied science faststart high fidelity pcr system 1 is the product of choice for multiplex pcr. This protein mediates proofreading activity, but has no polymerase activity itself. Download a pdf containing pricing for our full product list. Fidelity pcr system, or instead of single reagents, for nearly all pcr. The faststart high fidelity pcr system an enzyme blend for hot start.
Chapter 4 general pcr methods michigan state university. Faststart taq dna polymerase is the best pcr enzyme for most uptodate standard. High fidelity pcr products your most demanding applications, such as cloning and expression, sequencing, and genotyping require highly accurate results. The faststart high fidelity pcr system is the product of choice for multiplex pcr. Superscript iii rt platinum taq high fidelity enzyme mix and 2x reaction mix. This protein mediates proofreading activity, but carries no polymerase activity.
Expand high fidelityplus pcr system preventing carryover, protocol m page 1. The system uses a mixture of superscript iii reverse. The supplied high fidelity pcr master mix is sufficient for 200 amplifi cation reactions of 50. Demonstration of the expand tm pcr systems greater. The phusion high fidelity pcr kit contains a sufficient supply of phusion high fidelity dna polymerase, phusion hf and gc buffers, deoxynucleotides, magnesium chloride, dmso, and dna size standard to perform 50 reactions small or 200 reactions large.
Faststart taq dna polymerase is a versatile enzyme that can be used in a wide variety of applications and on multiple instrument platforms. After 30 pcr cycles, the 10 kb fragment must be clearly visible after agarosegel electrophoresis and ethidium bromide staining. The superscript iii onestep rtpcr system with platinum taq high fidelity is designed for convenient endpoint detection and analysis of rna molecules by onestep rtpcr. Faststart high fidelity enzyme blend with additional readytouse 10 mm pcr grade nucleotide mix. Expand high fidelity pcr system is composed of a special enzyme mix containing. The aim of this study was to determine the efficiency of restorase, a novel dna polymerase blend that is known to repair damaged dna and the faststart high fidelity pcr system enzymes, on degraded forensic bovine samples using pcrbased methodology.
Pcr product size, primer sequences and annealing temperatures are shown in table 3. The phusion highfidelity pcr kit contains a sufficient supply of phusion highfidelity dna polymerase, phusion hf and gc buffers, deoxynucleotides, magnesium chloride, dmso, and. The accuscript high fidelity rtpcr system can amplify cdna of 0. Hot start taq dna polymerase for highly specific and sensitive amplification using pcr. Faststart high fidelity pcr system comprises thermostable proofreading dna polymerase, recommended for challenging pcrs. This enzyme mixture transcribes more accurately up to fourfold higher fidelity than taq dna polymerase alone and is better able to amplify sequences with high 40 60% gc content. Taq dna polymerase is designed for the sensitive, reproducible, endpoint. Pcr using nebnext high fidelity 2x pcr master mix m0541 pcr using q5 hot start high fidelity dna polymerase m0493 protocol for q5 hot start high fidelity 2x master mix. The enzyme dntpack comprises the faststart high fidelity pcr system and a readytouse solution of pcrgrade nucleotides.
Superscript iii onestep rtpcr system with platinum taq high. Also, there are modified versions of taq that include proofreading activity, such as faststart high fidelity pcr system and takara ex taq polymerase. Fulllength lncrnahit cdna was amplified using the faststart high fidelity pcr system roche applied science and nmumg genomic dna as a template. Demonstration of the expand tm pcr systems greater fidelity. Expand high fidelity plus pcr system from roche applied.
Tissue culture and enzyme freezer supply tcefs 2016. Superscriptz iii onestep rtpcr system with platinum. Thermo scientific phusion highfidelity dna polymerases have established a new standard for highfidelity pcr. Cloning primers contained ecori sites for inserting cdna into the pcdna3. The master mix contains an inert, nontoxic tracking dye to visualize mixing. Please see the online application note stepbystep protocol for multiplex pcr with the faststart high. The system uses a mixture of superscript iii reverse transcriptase and platinum taq high fidelity dna polymerase for enhanced rtpcr yields and fidelity, as well as the detection of longer templates. Tissue culture and enzyme freezer supply tcefs 2016 roche. The faststart high fidelity pcr system is an enzyme blend of chemically modified taq dna polymerase faststart taq dna polymerase and a thermostable via chemical modification proofreading protein. Fidelity pcr system shows a threefold increase compared to taq dna polymerase. For most of the experiments with this master mix, only template, primers and h 2 o will be needed. Tissue culture and enzyme freezer supply tcefs 2015. Ive used both, and had great success with faststart especially.
Applications the faststart high fidelity pcr system can be. Find safety data sheets, certificates of analysis and method sheets instructions for use in the roche support portal elabdoc in case of questions please. Faststart high fidelity pcr system from roche applied science. Hot start protocols have improved specificity, sensitivity, and pcr yield. For further processing on its own or in a mixture as part of an ivd method and under controlled conditions only acc. Highfidelity pcr products your most demanding applications, such as cloning and expression, sequencing, and genotyping require highly accurate results. Effect of the enzyme and pcr conditions on the quality of. Link roche applied science pcr application manual 3rd edition. The faststart high fidelity pcr system an optimized.
Biotaq bioline, london, uk, faststart high fidelity pcr system roche, mannheim, germany, amplitaq gold applied biosystems, warrington, uk, hotstartaq dna polymerase qiagen, hilden, gernamy, phusion high fidelity dna polymerase. Description high fidelity pcr enzyme mix is a unique blend of taq dna polymerase and a thermostable dna. Due to this unique fusion technique, phusion dna polymerases generate pcr products with accuracy and speed. Biotaq bioline, london, uk, faststart high fidelity pcr system roche, mannheim, germany. Tissue culture and enzyme freezer supply tcefs 2015 roche. Jan 27, 2015 a range of high fidelity, regular, economy and premium taq polymerase enzymes were selected. Each cycle involves three steps, which are described in detail above. Long noncoding rnas lncrna regulated by transforming.
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