The applied biosystems amplitaq gold 360 dna polymerase, when used with improved amplitaq gold 360 buffer, and the optional 360 gc enhancer, amplifies a vast range of dna sequence contexts. Abi prism and its design, applied biosystems, and microamp are registered. The amplitaq gold 360 master mix can be thawed once, then stored at 2 to 8 c. Techniques for minimizing the effects of pcr inhibitors in. Amplitaq gold 360 master mix protocol vii how to use this guide purpose of this guide the amplitaq gold 360 master mix protocol provides all the information you need to perform pcr over a wide range of dna templates, including some of the most challenging gcrich sequences. View online or download applied biosystems geneamp pcr system 9700 user manual. The multiplex pcr method for the detection of alloiococcus otitidis, haemophilus influenzae, moraxella catarrhalis, and streptococcus pneumoniae p. Applied biosystems simpliamp thermal cycler user manual pdf.
Amplitaq gold 360 dna polymerase delivers 360 coverage for a. Amplitaq gold dna polymerase with buffer ii and mgcl 2. The pcr process is a simple and powerful method, 5 invented by k. Two user attention words appear in applied biosystems user documentation. L with 10x amplitaq gold 360 pcr buffer, 25mm m2cl 2, and 360 gc enhancer. Applied biosystems amplitaq gold 360 dna polymerase. Protocol pu no man000 ev a0 amplitaq gold dna polymerase. Amplitaq gold dna polymerase is a chemically modified form of amplitaq dna polymerase requiring thermal activation. Clinically applicable multiplex pcr for four middle ear. Ampflstr identifiler pcr amplification kit user guide. The amplitaq gold dna polymerase, ld, 250 units, pn 4338856 reagent kit the amplitaq gold dna polymerase, ld, units, pn 4338857 reagent kit. Safety alert words also appear in user documentation. Page 71 appendix e predefined run methods location of power point and ports on the instrument amplitaq gold 360 method stage reaction cover cyclestep volume temp. With the wide template coverage, optimization of pcr reaction conditions, is virtually eliminated with the use of amplitaq gold 360 master mix.
Forensic biology protocols for forensic str analysisamplification using the yfilertm system status. Abi prism 7000 sequence detection systems relative. A portable document format pdf version of this guide is available on the veriti. Pdf advantages of a new taq dna polymerase in multiplex. Amplitaq gold 360 dna polymerase for superior performance also available is amplitaq gold 360 dna polymerase for even better pcr performance. Ampflstr identifiler and the amplitaq gold fast pcr master mix. Amplitaq gold is derived from recombinant, thermostable, 94 kda dna polymerase, encoded by a modified form of the thermus aquaticus dna polymerase gene and is available with 3 different buffers, which provide preferred ph and ionic strength for pcr amplif\ ication reactions. When used with the enhanced 360 gold buffer, this ultrapure enzyme, in addition to its hotstart capabilities, reduces false. Note see the interchangeable sample block module users manual for part. To verify your client connection to the database, you need a valid. Validation of the ampflstr sgm plus system for use in. Samples are injected for 5 seconds at 15,000 v and separate at 15,000 v for 21 minutes with run temperature of. Higher specificity, higher yield amplitaq gold dna polymerase can be activated partially or completely in a prepcr heat step, or can be allowed to activate slowly in a timereleased manner during the denaturation steps of thermal cycling.
Amplitaq gold 360 master mix thermo fisher scientific. In our current study, which involved pcr amplification using amplitaq gold fast pcr master mix, up, and a standard pcr thermal cycler rather than the fast pcr thermal cycler, we succeeded in allele typing while reducing pcr running times by half to approximately 90min. Provides information that may be of interest or help but is not critical to the use of. Results with amplitaq gold dna polymerase were more sensitive and specific compared with amplitaq dna polymerase, with and without manual hotstart procedure. Allele typing proved successful with all these methods, although amplification times were reduced by to 23 to approximately one to two hours. Amplitaq gold 360 master mix contains amplitaq gold 360 dna polymerase and optional 360 gc enhancer. A site preparation and safety guide is a separate document sent to all. Analyze samples as described in the users manual for the abi prism 3100 or. The following user attention words appear in applied biosystems user documentation. Chapters in the ampflstr profiler plus pcr amplification kit users.
User guide dna fragment analysis by capillary electrophoresis publication number 4474504 rev. Each word implies a particular level of observation or action as described below. Applied biosystems 7900ht fast realtime pcr system and. Amplitaq gold dna polymerase, ld hot start, strong finish protocol.
The purchase price of the ampflstr identifiler pcr amplification kit pn 4322288 includes limited, nontransferable rights under the runningroyalty component to use only this amount of that product to practice the polymerase. Designed for convenience, the amplitaq gold 360 pcr master mix scales to various reaction volumes for greater application and format. The pcr process is a simple and powerful method, 5. Applied biosystems 75007500 fast thermo fisher scientific. Papillocheck manual as pdf file portable download format on cd. If you are familiar with the instrument and its operation, proceed as follows. The taqman gold rtpcr kit contains multiscribe reverse transcriptase for first strand cdna synthesis, and thermal stable amplitaq gold dna polymerase for second strand cdna synthesis and dna amplification.
Applied biosystems amplitaq gold dna polymerase with buffer. Reading of the ampflstr profiler pcr amplification kit user manual is strongly recommended prior to. In this chapter the following topics are covered in this chapter. Prinz m 2003 advantages and disadvantages of yshort tandem repeat testing in forensic casework.
Storage when you receive the amplitaq gold 360 master mix, store at. Four safety alert words appear in applied biosystems user documentation at points. Str identifiler pcr amplification kit user guide for use with. User attention words throughout the abi p rism sequence detection system users manual, four kinds of information are set off from the regular text. Amplitaq gold 360 pcr master mix contains everything required for successful pcr amplification in one convenient packageall components are premixed and premeasured. For manual cycle sequencing of dna, use the amplicycle sequencing kit n8080175 featuring amplitaq dna polymerase, cs. This manual provides procedures for operating the abi p rism 7700 sequence detector, describes instrument and software features, and presents some theory and guidelines to operation. The master mix is supplied at 2x the recommended usage concentration for easy dilution when adding template and primers. The amplitaq gold pcr with bsa was tested for its effectiveness at minimizing the effects of pcr inhibition using peruvian amphibian skin swabs that were collected in 2011. Amplitaq gold 360 dna polymerase delivers 360 coverage for a full range of targets. It is readily absorbed through the skin and is a combustable liquid and vapor keep away from heat and flame. Amplitaq gold is derived from a recombinant, thermostable, 94 kda dna polymerase encoded by a modified form of the thermus aquaticus dna polymerase gene which has been inserted into an escherichia coli host. When using amplitaq gold enzyme, you can introduce hot start pcr and time release pcr.
The ampflstr profiler plus pcr amplification kit users manual pn 4303501 should be referred to for use with the ampflstr profiler plus id kit. Short tandem repeat analysis for human identity testing. At a previous conference, we reported on fast pcr cycling for ampflstr identifiler by three methods. Pdf advantages of a new taq dna polymerase in multiplex pcr. The results were compared with that of an amplitaq gold pcr using the same samples. Amplitaq gold 360 dna polymerase is supplied at 5u. Introduction and safety 11 introduction and safety 1 overview about this chapter this chapter provides information to help you safely operate the geneamp pcr system 9700. Applied biosystems geneamp pcr system 9700 pdf user manuals. View and download applied biosystems simpliamp thermal cycler user manual online. Automated chemical hotstart enzyme for increased specificity, sensitivity, and yield timereleased thermal activation improves sensitivity in low copy number. Applied biosystems veriti thermal cycler quick reference card provides a summary of information about the thermal cycler, its installation, and accessories. Amplitaq gold 360 dna polymerase provides the same hotstart specificity as amplitaq gold dna polymerase. Amplitaq gold 360 master mix contains amplitaq gold.
Powerplex 16 system technical manualpdf promega corporation. All necessary materials are provided to amplify str regions of human genomic dna. We have 3 applied biosystems geneamp pcr system 9700 manuals available for free pdf download. Fast pcr amplification of ampflstr yfiler forensic. We would like to show you a description here but the site wont allow us. Dye, amplitaq gold dna polymerase, dntps with dutp, passive reference, and optimized buffer components. Simpliamp thermal cycler bicycle accessories pdf manual download. Quantstudio 6 and 7 flex realtime pcr system software. Sybr green pcr master mix and rtpcr reagents protocol. All other trademarks are the sole property of their respective owners. The main ingredient of the master mix is amplitaq gold 360 dna polymerase but it also contains the worldclass 360 gc enhancer which can be added optionally for high gccontent templates. Applied biosystems 75007500 fast realtime pcr system getting started guide for genotyping experiments v preface how to use this guide about the system documentation the guides listed below are shipped with the applied biosystems 75007500 fast realtime pcr system 75007500 fast system.
Storage buffer enzyme is supplied in 20mm tris, ph 9. Amplitaq gold dna polymerase, ld hot start, strong finish protocol 15 materials and equipment two kits available amplitaq gold dna polymerase, ld is available in two kit configurations. This guide uses conventions and terminology that assume a working knowledge of the microsoft windows operating system, the internet, and internetbased browsers. Firsttime users of the quantstudio 6 and 7 flex realtime pcr system. Sybr green may cause eye, skin, and respiratory tract irritation. Taqman universal pcr master mix harvard university. Pullout chapters this user manual contains individually bound chapters chapter 2 through chapter 6 that the user can pull out of the binder to use in the lab.
Sybr green pcr master mix and rtpcr reagents protocol pn. The ampflstr profiler plus pcr amplification kit users manual pn 4303501. For the current study, we performed pcr amplification with ampflstr yfiler using amplitaq gold fast pcr master mix, upx2. Typically, msi analysis involves comparing allelic profiles of microsatellite markers generated by amplification of dna from matching normal and test samples, which may be mismatchrepair mmr deficient. A hightemperature incubation step is required to activate amplitaq gold dna polymerase, which ensures that the active enzyme is generated only at temperatures in which the dna is fully denatured and when the primers are not annealed. Applied biosystems amplitaq gold dna polymerase with. Amplitaq gold fast pcr master mix, up 2x protocol, applied biosystems. Amplitaq gold dna polymerase with buffer ii thermo fisher. Powerplexr fusion 6c system for use on the applied. With or without a limited upfront heat activation step, active enzyme is released slowly during thermal. Ampflstr identifiler pcr amplification kit users manual pcr setup 1. Compared to amplitaq gold dna polymerase, amplitaq gold 360 dna polymerase is purified by an additional proprietary separation process to eliminate contaminating bacterial dna sequences from the enzyme preparation.
Because amplitaq gold dna polymerase is a chemical hotstart enzyme, there is no worry of biological contamination. Synthesis of 1st cdna strand oligo dt or random hexamer 3. The amplitaq gold 360 dna polymerase protocol provides all the. Google scholar the pcr cycling parameters were prepcrdenaturation at 95 c for 10 min, followed by 10 cycles at 94 c at 30 s 50%, 61 c for 1 min, 72 c for 30 s, 20 cycles of 94 c for 5 s 50%, 61 c for 1 min, 68 c for 10 s, and 60 c for 30 min. Optimized for broadest range of targetsfrom everyday to challenging.
Amplitaq gold is a thermostable dna polymerase designed to make. A subset of these samples that were either bdpositive, bdnegative. Amplitaq gold 360 master mix was designed for 360 coverage of a full range of targets. Applied biosystems amplitaq gold dna polymerase with gold. Amplitaq gold dna polymerase, ld protocol thermo fisher. The calibrate function is also available in the control console. To detect falsenegative results, an internal amplification. A portable document format pdf version of this guide is. This manual contains a protocol for use of the powerplex fusion 6c system with the geneamp pcr system 9700, veriti 96well thermal cycler and proflex pcr system in addition to protocols to separate amplified products and detect separated material on the. Applied biosystems 7900ht fast realtime pcr system and sds. Amplitaq gold dna polymerase the amplitaq gold enzyme is a thermal stable dna polymerase. For more information, see safety alert words on page vii.
At isfg2007, an earlier conference, we presented reports on two fast pcr cycling methods using ampflstr identifiler. Amplitaq gold 360 master mix 1 ml, 360 gc enhancer on receipt, store at. Dynamic mechanical analysis accurate, precise, versatile dma measurements. Applied biosystems amplitaq gold 360 dna polymerase 100. Activity is restored after the denaturation step in pcr cycling at 94c, providing an automatic hot start and offering increased sensitivity, specificity, and yield, while allowing assembly of reactions at room temperature. Fast pcr amplification of ampflstr yfiler forensic science. Amplitaq gold 360 master mix is available in the following packaging. Onestep rtpcr onestep rtpcr uses a single buffer that enables rt and pcr amplification to occur without interruption. Relative quantification getting started guide for the 73007500 system iii 3. It is designed to help you quickly learn to use the veriti thermal cycler.
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