PyroMark Q24 Advanced

For advanced methylation and mutation quantification in long sequence runs using Pyrosequencing
  • Advanced technology, software, and chemistry for long sequence runs
  • Quantitative methylation analysis at consecutive CpG or CpN sites
  • Improved quantification of sequence variations at any sequence position
  • Easy and improved base calling functionality
  • Assay versatility on the same instrument and in the same run
PyroMark Q24 Advanced has improved Pyrosequencing technology to provide even better real-time sequence-based detection and quantification than before. PyroMark Q24 Advanced features advanced technology, software, and chemistry, and is highly suited for analyzing any kind of sequence variation, particularly DNA methylation at CpG or CpN sites, complex mutations, or for de novo sequencing applications such as microbial typing.
产品 Product no. 货号 目录价:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
产品 货号 目录价:
PyroMark Q24 Advanced System
Instrument and software for advanced Pyrosequencing analysis: includes installation, training, and 1-year warranty on parts and labor
9002270
询价
PyroMark Q24 Advanced Priority Package
Instrument and software for advanced Pyrosequencing analysis: includes Priority Package with installation, training, 2-year warranty on parts and labor, and 2 preventive maintenance visits
9002272
询价
PyroMark Q24 Advanced Priority Package Plus
Instrument and software for advanced Pyrosequencing analysis: includes Priority Package Plus with installation, training, 3-year warranty on parts and labor, and 3 preventive maintenance visits
9002273
询价
PyroMark Q24 Advanced Software
Analysis software for upgrading PyroMark Q24 systems
9022779
询价

PyroMark Q24 Advanced and PyroMark Q24 Advanced Software are intended to be used only in combination with QIAGEN kits indicated for use with the PyroMark Q24 Advanced for the applications described in the kit handbooks. If PyroMark Q24 Advanced and PyroMark Q24 Advanced Software are used with other than QIAGEN kits, it is the user's responsibility to validate the performance of such product combination for any particular application.
在长片段测序中分析16个CpG位点。|焦磷酸测序原理 — 第1步。|焦磷酸测序原理 — 第2步。|焦磷酸测序原理 — 第3步。|焦磷酸测序原理 — 第4步。|焦磷酸测序原理 — 第5步。|对均聚物进行更好的甲基化定量分析。|长片段从头测序。|对长片段进行定量突变分析。|工作流程解决方案。|完全整合的系统。|便利的数据管理。|高效的模板制备。|
PyroMark Q24 Advanced提高了测序长度和较靠后位点的甲基化分析可靠性。图中展示了PyroMark Q24 Advanced进行CpG分析时对135个核苷酸的序列中16个不同CpG位点进行的准确分析。如使用PyroMark Q24则需分别进行3次反应。|扩增一DNA片段,将该片段生物素化,作为焦磷酸测序的模板。变性后,生物素化的单链PCR扩增子分离,与测序引物杂交。杂交的引物和单链模板,与DNA聚合酶、ATP硫酸化酶、萤光素酶、三磷酸腺苷双磷酸酶,以及腺苷5'磷酸化硫酸(APS)和萤光素混合进行孵育。|将第一个dNTP加入反应液中。在DNA聚合酶作用下,dNTP与测序引物结合,按照碱基互补原则,引物与模板结合。每次结合都会释放出PPi,其释放量与结合的核苷酸呈等摩尔关系。|在腺苷5'磷酸化硫酸(APS)存在下,ATP硫酸化酶将PPi转化为ATP。ATP驱动荧光素在荧光素酶介导下转化为氧化萤光素,产生可见光,光的量与ATP量成正比。荧光素酶催化的反应中产生的光由CCD传感器检测,在输出的原始数据中(Pyrogram)表现为峰值。每个峰的高度(光信号)与结合的核苷酸的数量成比例。|腺苷三磷酸双磷酸酶是一种核苷酸降解酶,不断降解未结合的核苷酸和ATP。降解完成时,另一个核苷酸被添加上。|按照顺序添加dNTP。应当注意:α-硫代三磷酸脱氧腺苷(dATPαS)被用作天然脱氧腺苷三磷酸(dATP)的替代品,因为它能被DNA聚合酶高效利用,但不会被荧光素酶识别。在处理过程中,互补的DNA链逐渐增长,核苷酸的顺序通过Pyrogram图中信号峰来确定。|对紧邻T均聚物之后的、或T均聚物之间的CpG位点进行甲基化分析十分具有挑战性。甲基化程度由亚硫酸盐处理后,转换的C碱基与未转换的C碱基的比例决定,因为C到T的转换往往导致长T均聚物在扩增子中伸展,这种情况经常发生。 PyroMark Q24 Advanced能够对紧邻T均聚物之后的、或T均聚物之间的CpG位点进行准确的定量分析。图中例子显示了对8个T碱基内的CpG位点的分析。|序列读长是对未知序列进行分析的关键因素,通常仅可对40–80各碱基进行可靠分析。PyroMark Q24 Advanced利用改善的试剂和算法,显著提高了序列读长,并提供更高的准确性。使用PyroMark Q24(上方)和PyroMark Q24 Advanced(下方)和相同的试剂进行从头测序。蓝色柱形表示可靠检测到的碱基;黄色表示可能正确检测的碱基,但用户应检查确定;红色表示不可靠的检测结果。|由于突变和单核苷酸多态性(SNP)很少在直接邻近位置发生,因此常用的焦磷酸测序试剂在分析一个以上的突变位点时,可能需要不同试剂。PyroMark Q24 Advanced利用新型试剂,具有更长的序列读长,能够在一次运行中可靠分析多个突变。图中例子显示了10:90的EGFR野生型和突变型混合物的分析。即使在分析了60个核苷酸之后,分析仍是准确的。|PyroMark Q24可简单、高效的整合入焦磷酸测序工作流程。每一步,从引物设计到PCR扩增和测序模板制备,都由专为焦磷酸测序设计的软件、试剂盒、试剂和样本制备仪器提供高品质支持。|PyroMark Q24具有较小的体积,但能够胜任快速处理24个样本测序的各个步骤。只需将样本和试剂上样,上传实验文件,测序实验即可自动进行。PyroMark Q24可准确的将试剂和核苷酸加入每个孔中。反应时会发出光信号,由24个CCD传感器(每孔一个)检测,因此可避免信号交叠。|PyroMark Q24可单独摆放,因此可便利的放置于实验室任何位置。数据存储于仪器的硬盘中,可在实验进行时在显示屏上查看数据。此外,所有文件存储在U盘中,用户可便利的在安装有PyroMark Q24 Software的电脑上分析数据。|PyroMark Q24 Vacuum Workstation可将PCR产物转化为单链DNA用于焦磷酸测序。将PCR扩增子加入多重优化的溶液中进行变性,然后洗涤获得DNA。对24个样本平行进行这一流程,仅需花费几分钟。|
性能
Longer Pyrosequencing runs and improved accuracy
PyroMark Q24 Advanced features improved chemistry and instrument operation algorithms that significantly increase the assay read length and accuracy of the base calling functionality, enabling easy analysis of long de novo sequencing runs. Assay read length was previously limited by background peaks and reduced light signals in the sequencing reaction. The updated chemistry and algorithms of PyroMark Q24 Advanced reduce these background peaks, thereby increasing read length and reliability. Depending on the sequence to be analyzed, highly accurate read lengths of 140 or more bases can be obtained in just a single PyroMark Q24 Advanced reaction (see figure Long de novo sequencing runs).

Improved methylation analysis at any position
PyroMark Q24 Advanced enables improved methylation quantification in long sequence runs at any sequence position. Previously, analysis of methylation sites further away from the sequencing primer could be uncertain, but now with longer read lengths and higher accuracy, methylation quantification is highly reliable throughout the entire sequencing run (see figure Analysis of 16 CpG sites in a long sequence run).

Improved sequencing accuracy in homopolymers
Bisulfite conversion in DNA methylation analysis leads to frequent poly T stretches in the nucleotide sequence, and analysis of CpG sites directly after such T homopolymers has previously been challenging due to uncertain quantification of the light signal at these sites. The increased accuracy of PyroMark Q24 Advanced enables reliable quantification of CpG methylation behind and even within a stretch of 8 T nucleotides (see figure Improved methylation quantification in homopolymers).

Analysis of mutations over long sequences
PyroMark Q24 Advanced also provides reliable quantification of multiple polymorphisms in a single assay. Since single nucleotide polymorphisms (SNPs) and other mutations are often not located close to one another, traditional Pyrosequencing chemistry usually requires separate assays for each mutation site to be analyzed. The new chemistry of PyroMark Q24 Advanced allows much longer runs, enabling reliable analysis of more than one mutation or SNP in the same run (see figure Quantitative mutation analysis in long sequence runs).
原理
Pyrosequencing technology, which is based on the principle of sequencing by synthesis, provides quantitative data in sequence context within minutes. PyroMark Q24 Advanced is a fully integrated system that provides real-time sequence information with high accuracy and long read lengths, making it highly suitable for analysis of complex mutations, epigenetic research, resistance typing, and microbial identification. The system comprises the PyroMark Q24 instrument (running firmware 3.0 or higher), PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents/PyroMark Q24 Advanced CpG Reagents, PyroMark Q24 Vacuum Workstation, and PyroMark Control Oligo (see table). Sample preparation solutions are also supplied to enable preparation of single-stranded DNA using the PyroMark Q24 Vacuum Workstation.

Component                            Description
PyroMark Q24 Sequencing instrument for quantitative sequence analysis
PyroMark Q24 Advanced Software Analysis software with 4 analysis modes (AQ: allele quantification; SNP: genotyping; CpG: methylation at CpG or CpN sites; and SEQ: base-calling of unknown sequences)
PyroMark Q24 Advanced Reagents Enzymes, substrates, nucleotides, and buffers. Compatible only with assays set up and analyzed with PyroMark Q24 Advanced Software. 
PyroMark Q24 Advanced CpG Reagents  Enzymes, substrates, nucleotides, and buffers for assays requiring longer sequence read lengths. Compatible only with assays set up and analyzed with PyroMark Q24 Advanced Software.
PyroMark Q24 Vacuum Workstation Workstation for preparing up to 24 samples in parallel from PCR product to single-stranded template 
PyroMark Q24 Control Oligo Control for verification of proper installation and operation of the system
PyroMark Q24 Advanced system.
Steps of the Pyrosequencing reaction:
Step 1: A DNA segment is amplified, and the strand to serve as the Pyrosequencing template is biotinylated. After denaturation, the biotinylated single-stranded PCR amplicon is isolated and allowed to hybridize with a sequencing primer. The hybridized primer and single-stranded template are incubated with the enzymes DNA polymerase, ATP sulfurylase, luciferase, and apyrase, as well as the substrates adenosine 5' phosphosulfate (APS) and luciferin (see figure "Principle of Pyrosequencing — step 1").

Step 2:
The first deoxribonucleotide triphosphate (dNTP) is added to the reaction. DNA polymerase catalyzes the addition of the dNTP to the squencing primer, if it is complementary to the base in the template strand. Each incorporation event is accompanied by release of pyrophosphate (PPi), in a quantity equimolar to the amount of incorporated nucleotide (see figure "Principle of Pyrosequencing — step 2").

Step 3: ATP sulfurylase converts PPi to ATP in the presence of adenosine 5' phosphosulfate (APS). This ATP drives the luciferase-mediated conversion of luciferin to oxyluciferin that generates visible light in amounts that are proportional to the amount of ATP. The light produced in the luciferase-catalyzed reaction is detected by CCD sensors and seen as a peak in the raw data output (Pyrogram). The height of each peak (light signal) is proportional to the number of nucleotides incorporated (see figure "Principle of Pyrosequencing — step 3").

Step 4:
Apyrase, a nucleotide-degrading enzyme, continuously degrades unincorporated nucleotides and ATP. When degradation is complete, another nucleotide is added (see figure "Principle of Pyrosequencing — step 4").

Step 5: Addition of dNTPs is performed sequentially. It should be noted that deoxyadenosine alpha-thio triphosphate (dATPαS) is used as a substitute for the natural deoxyadenosine triphosphate (dATP), since it is efficiently used by the DNA polymerase, but not recognized by the luciferase. As the process continues, the complementary DNA strand is elongated, and the nucleotide sequence is determined from the signal peaks in the Pyrogram trace (see figure "Principle of Pyrosequencing — step 5").
 
Streamlined workflow — from sample to result
The versatile PyroMark Q24 Advanced seamlessly integrates into epigenetic and genetic analysis workflows, and complements QIAGEN's advanced technologies for sample preparation, bisulfite conversion, and PCR amplification. The highly reliable instrument enables sequence-based detection and quantification of methylation at CpG or CpN sites, as well mutations. The streamlined workflow means that results can be achieved faster.
操作流程
Fast and easy sample preparation 
From PCR product to single-stranded template ready for sequencing — up to 24 samples can be prepared in parallel using the PyroMark Q24 Vacuum Workstation, in less than 15 minutes. The workstation ensures easy handling, and the actual "hands-on time" is less than 5 minutes.

Prior to Pyrosequencing, a biotinylated PCR product is generated. This biotinylated PCR product is bound to Streptavidin-coated Sepharose beads, and the beads are captured with the Vacuum Tool on the Vacuum Workstation, where they are thoroughly washed and subsequently denatured, generating single-stranded DNA suitable for Pyrosequencing. This template DNA is released into the Pyrosequencing reaction plate containing the sequencing primer, and after primer annealing, the plate is placed into the PyroMark instrument. PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents contain the enzymes, nucleotides, and substrate for the Pyrosequencing reaction; these are pipetted into the dispensing cartridge, according to the volumes provided by the software, and are also placed into the instrument for the Pyrosequencing run.
应用
Pyrosequencing is becoming increasingly important for research applications in a variety of disciplines. Whether examining drug-resistance development in pathogens, the role of epigenetic DNA methylation in gene expression regulation, genetic markers for specific phenotypes in livestock, or polymorphisms in forensic samples of mitochondrial DNA, the PyroMark Q24 Advanced enables powerful and versatile analysis of genetic and epigenetic variation. In addition, because Pyrosequencing integrates sequence detection and quantification, the enhanced analysis resolution can lead to new discoveries.

Product comparison
  PyroMark Q24 Advanced PyroMark Q24
Throughput 1–24 samples 1–24 samples
Running volume 25 µl 25 µl 
PCR requirements 5–10 µl
(~0.5–3 pmol of product)
5–10 µl
(~0.5–3 pmol of product)            
Read length (assay- and
sequence-dependent)
10–140 bp or more 10–80 bp
Application software PyroMark Q24 Advanced SW
(requires firmware 1.5.6903 or higher)
PyroMark Q24 SW 2.0
Software functionality SEQ (de novo sequencing)
CpG/CpN methylation
SNP
AQ
SQA (de novo sequencing)
CpG methylation
AQ/SNP 
 
Main applications Complex mutation analysis
Epigenetics (CpG and CpN analysis)
Resistance typing and microbial ID  
Mutation analysis
Resistance typing 
 
Compatible reagents PyroMark Q24 Advanced Reagents
PyroMark Q24 Advanced CpG Reagents  
PyroMark Q24 Gold Reagents 
Sensitivity 2% mutation
98% wt  
2% mutation
98% wt  

软件
Easy-to-use PyroMark Q24 Advanced Software
PyroMark Q24 Advanced Software, installed on a PC, enables comprehensive analysis of your results. The software contains 4 analysis modes — AQ, SNP, CpG, and SEQ. The AQ mode can be used for a variety of quantification studies of mutations such as SNPs and InDels. It is suitable for analyzing single and multivariable positions, as well as di-, tri- , and tetra- allelic mutations. The SNP mode provides genotype analysis of SNPs and InDels. The CpG mode enables methylation analysis of single or multiple CpG or CpN sites and provides a built-in control for the bisulfite treatment. The SEQ mode is used for base-calling of unknown sequences.

The PyroMark Q24 Advanced Software is user-friendly and intuitive and provides convenient and improved tools for run analysis. If a problem occurs during the run, or if the system detects an inconsistency with an assay, the software provides specific warning information for each individual well (see Warning information and recommendations). A “Warning Info” button gives access to additional information about the warning along with recommendations for troubleshooting and preventing its occurrence in subsequent assays.

Flexible and simple Pyrosequencing assay design using PyroMark Assay Design Software
PyroMark Assay Design Software 2.0 ensures easy design of PCR and sequencing primers for Pyrosequencing analyses. The assays are optimized for use with all PyroMark instruments.
特点
参数
海拔 Up to 2000 m (6500 ft)
应用 Methylation analysis, allele quantification, genotyping, sequence analysis
化学试剂抗性 pH 4 to pH 9, common detergents, 0.5 M sodium hydroxide, ethanol
连接 One USB port (2.0)
湿度 Relative humidity of 20–90% (noncondensing)
仪器尺寸 420 x 390 x 525 mm (16.5 x 15.4 x 20.7 in)
配合仪器使用的试剂 PyroMark Q24 Tests
运行温度 15–32°C (59–90°F)
过电压保护类别 II
操作地点 For indoor use only
污染水平 2
功率 100–240 V AC, 47–63 Hz, 1.1–0.45 A (grounded); from external power supply to the instrument : 12 VDC and 24 VDC nominal
处理温度 28°C (82.4°F) ± 1%
处理时间 Depends on the number of dispensations (20 dispensations take 24 minutes)
每次处理样本量;通量 1–24
软件 PyroMark Q24 Software 2.0 (to be installed on PC)
技术 Pyrosequencing
重量 27.5 kg (60.6 lb)

您无权限查看此资源

Download Files
14
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0015
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0014
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0003
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0013
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0005
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0008
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0006
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0011
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0004
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0009
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0007
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0012
显示更多
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0010
显示更多
Files for upgrading existing PyroMark Q24 instruments to firmware 1.5.6903
显示更多
Operating Software
2
PyroMark Q24 Advanced Software version 3.0.1 is compatible with Windows 7 and Windows 10 (64 bit) operating systems. This software may only be downloaded by registered users with a valid PyroMark Q24 Advanced Software license and registered PyroMark Q24 Advanced instrument. If you do not have a valid software license, contact your QIAGEN sales representative.
显示更多
Files for upgrading existing PyroMark Q24 instruments to firmware 1.5.6903
显示更多
图片
Analysis of 16 CpG sites in a long sequence run
在长片段测序中分析16个CpG位点。
PyroMark Q24 Advanced提高了测序长度和较靠后位点的甲基化分析可靠性。图中展示了PyroMark Q24 Advanced进行CpG分析时对135个核苷酸的序列中16个不同CpG位点进行的准确分析。如使用PyroMark Q24则需分别进行3次反应。
Principle of Pyrosequencing – Step 1
焦磷酸测序原理 — 第1步。
扩增一DNA片段,将该片段生物素化,作为焦磷酸测序的模板。变性后,生物素化的单链PCR扩增子分离,与测序引物杂交。杂交的引物和单链模板,与DNA聚合酶、ATP硫酸化酶、萤光素酶、三磷酸腺苷双磷酸酶,以及腺苷5'磷酸化硫酸(APS)和萤光素混合进行孵育。
Principle of Pyrosequencing – Step 2
焦磷酸测序原理 — 第2步。
将第一个dNTP加入反应液中。在DNA聚合酶作用下,dNTP与测序引物结合,按照碱基互补原则,引物与模板结合。每次结合都会释放出PPi,其释放量与结合的核苷酸呈等摩尔关系。
Principle of Pyrosequencing – Step 3
焦磷酸测序原理 — 第3步。
在腺苷5'磷酸化硫酸(APS)存在下,ATP硫酸化酶将PPi转化为ATP。ATP驱动荧光素在荧光素酶介导下转化为氧化萤光素,产生可见光,光的量与ATP量成正比。荧光素酶催化的反应中产生的光由CCD传感器检测,在输出的原始数据中(Pyrogram)表现为峰值。每个峰的高度(光信号)与结合的核苷酸的数量成比例。
Principle of Pyrosequencing – Step 4
焦磷酸测序原理 — 第4步。
腺苷三磷酸双磷酸酶是一种核苷酸降解酶,不断降解未结合的核苷酸和ATP。降解完成时,另一个核苷酸被添加上。
Principle of Pyrosequencing – Step 5
焦磷酸测序原理 — 第5步。
按照顺序添加dNTP。应当注意:α-硫代三磷酸脱氧腺苷(dATPαS)被用作天然脱氧腺苷三磷酸(dATP)的替代品,因为它能被DNA聚合酶高效利用,但不会被荧光素酶识别。在处理过程中,互补的DNA链逐渐增长,核苷酸的顺序通过Pyrogram图中信号峰来确定。
Improved methylation quantification in homopolymers
对均聚物进行更好的甲基化定量分析。
对紧邻T均聚物之后的、或T均聚物之间的CpG位点进行甲基化分析十分具有挑战性。甲基化程度由亚硫酸盐处理后,转换的C碱基与未转换的C碱基的比例决定,因为C到T的转换往往导致长T均聚物在扩增子中伸展,这种情况经常发生。 PyroMark Q24 Advanced能够对紧邻T均聚物之后的、或T均聚物之间的CpG位点进行准确的定量分析。图中例子显示了对8个T碱基内的CpG位点的分析。
Long de novo sequencing runs (sequence detail 95-140 bp) — PyroMark Q24 vs. PyroMark Q24 Advanced
长片段从头测序。
序列读长是对未知序列进行分析的关键因素,通常仅可对40–80各碱基进行可靠分析。PyroMark Q24 Advanced利用改善的试剂和算法,显著提高了序列读长,并提供更高的准确性。使用PyroMark Q24(上方)和PyroMark Q24 Advanced(下方)和相同的试剂进行从头测序。蓝色柱形表示可靠检测到的碱基;黄色表示可能正确检测的碱基,但用户应检查确定;红色表示不可靠的检测结果。
Quantitative mutation analysis in long sequence runs
对长片段进行定量突变分析。
由于突变和单核苷酸多态性(SNP)很少在直接邻近位置发生,因此常用的焦磷酸测序试剂在分析一个以上的突变位点时,可能需要不同试剂。PyroMark Q24 Advanced利用新型试剂,具有更长的序列读长,能够在一次运行中可靠分析多个突变。图中例子显示了10:90的EGFR野生型和突变型混合物的分析。即使在分析了60个核苷酸之后,分析仍是准确的。
工作流程解决方案。
PyroMark Q24可简单、高效的整合入焦磷酸测序工作流程。每一步,从引物设计到PCR扩增和测序模板制备,都由专为焦磷酸测序设计的软件、试剂盒、试剂和样本制备仪器提供高品质支持。
完全整合的系统。
PyroMark Q24具有较小的体积,但能够胜任快速处理24个样本测序的各个步骤。只需将样本和试剂上样,上传实验文件,测序实验即可自动进行。PyroMark Q24可准确的将试剂和核苷酸加入每个孔中。反应时会发出光信号,由24个CCD传感器(每孔一个)检测,因此可避免信号交叠。
便利的数据管理。
PyroMark Q24可单独摆放,因此可便利的放置于实验室任何位置。数据存储于仪器的硬盘中,可在实验进行时在显示屏上查看数据。此外,所有文件存储在U盘中,用户可便利的在安装有PyroMark Q24 Software的电脑上分析数据。
高效的模板制备。
PyroMark Q24 Vacuum Workstation可将PCR产物转化为单链DNA用于焦磷酸测序。将PCR扩增子加入多重优化的溶液中进行变性,然后洗涤获得DNA。对24个样本平行进行这一流程,仅需花费几分钟。