QuantiTect SYBR® Green RT-PCR Kit

使用SYBR Green I进行一步法定量RT-PCR,用于基因表达分析

  • 一步法RT-PCR具有高特异性
  • 可靠定量检测低丰度转录本
  • 准确定量检测各种起始量样本
  • 无需优化反应和循环条件

QuantiTect SYBR Green RT-PCR Kit使用SYBR Green I通过一步法real-time RT-PCR对RNA目标序列进行高特异性的定量检测。热启动和独特的实时定量PCR缓冲液确保了对RNA目标序列高特异性和灵敏度的real-time定量检测。dNTP混合液含有dUTP,可选择性进行UNG处理。QuantiTect SYBR Green RT-PCR Kit还含有优化的RT混合液,可对多种起始量的模板RNA进行高效、灵敏的逆转录。为便于使用,预混液可储存在2–8°C。

Product Product no. Cat. no. List price:
 
 
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.
Product Cat. no. List price:
QuantiTect SYBR Green RT-PCR Kit (200)
For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect SYBR Green RT-PCR Master Mix, 100 µl QuantiTect RT Mix, 2 x 2 ml RNase-Free Water
204243
Inquire
QuantiTect SYBR Green RT-PCR Kit (1000)
For 1000 x 50 µl reactions: 25 ml 2x QuantiTect SYBR Green RT-PCR Master Mix, 0.5 ml QuantiTect RT Mix, 20 ml RNase-Free Water
204245
Inquire
The QuantiTect SYBR® Green RT-PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
  • Main Image Navi
One-step RT-PCR.|Highly specific amplification.|Comparable results in one-step and two-step RT-PCR.|Specific primer annealing.|
The QuantiTect SYBR® Green RT-PCR Kit overcomes the need for optimization of reaction conditions, which can be tedious and time-consuming. Simply add primers and template to the ready-to-use RT-PCR master mix, and start the reaction on any real-time cycler.|In comparison with other DNA polymerases, only HotStarTaq DNA Polymerase in combination with a unique buffer specifically amplified a 497 bp fragment (from 50 copies of an HIV-pol-gene construct in a background of 1 µg human genomic DNA). M: Markers.|Total RNA (10 ng to 100 pg) or the equivalent amounts of cDNA from HeLa cells were analyzed using the QuantiTect Primer Assay for human MAPK14 and the indicated kits. Similar CT values were achieved in one-step and two-step RT-PCR.|A balanced combination of KCl and (NH4)2SO4 promotes specific annealing of primers to the PCR template. K+ binds to phosphate groups on double-stranded DNA, stabilizing primer annealing. NH4+ destabilizes weak hydrogen bonds between mismatched bases.|
Performance
QuantiTect SYBR Green RT-PCR Kit含有独特的逆转录酶,可高效、灵敏的进行cDNA合成在宽泛的范围内符合线性规律(参见"Comparable results in one-step and two-step RT-PCR")。HotStarTaq DNA Polymerase提供严格的热启动,进一步提高了PCR反应的特异性(参见"Highly specific amplification")。
Principle

QuantiTect SYBR Green RT-PCR Kits含有优化的即用型预混液,可使用SYBR Green I对RNA靶序列进行高特异性和灵敏度的real-time定量分析。预混液中的荧光染料SYBR Green I可对很多不同的靶序列进行分析,无需合成靶标特异性标记引物。独特的PCR缓冲液含有K+和NH4+离子,能促使特异性引物退火,获得高RT-PCR特异性和灵敏度(参见"Specific primer annealing")。此外,HotStarTaq DNA Polymerase具有严格的热启动,可防止非特异性产物的形成。

QuantiTect SYBR Green RT-PCR预混液含有dUTP,可在PCR前使用尿嘧啶DNA糖基化酶(UNG)预处理,保证之前反应残留的PCR产物不会影响后续的PCR反应。

2x QuantiTect SYBR Green RT-PCR Kit的组分
组分 特点 优势
HotStarTaq DNA Polymerase 95ºC孵育15分钟即可活化 室温下即可构建qPCR反应体系
QuantiTect SYBR Green RT-PCR Buffer NH4+和K+的平衡组合 特异性引物退火确保PCR结果可靠
dNTP mix 含有dUTP,会部分替代dTTP,可进行UNG处理 可选择进行UNG处理,以消除残余PCR产物的污染物
SYBR Green I染料 DNA双链结合时,生成强烈的荧光信号 高度灵敏的定量检测
ROX染料 在Applied Biosystems或Agilent的PCR仪上进行荧光信号校准 可在需要ROX染料的PCR仪上进行准确的定量检测。不会干扰real-time PCR反应
Omniscript和Sensiscript Reverse Transcriptases 对RNA具有高亲和性的酶 进行RNA逆转录,包括复杂二级结构的逆转录
Procedure

QuantiTect SYBR Green RT-PCR Kit无需进行繁琐耗时的反应条件优化。只需简单地将引物和模板加入到即用型RT-PCR预混液中,即可开始反应(参见"One-step RT-PCR")。使用操作手册中的实验方案可在任何real-time PCR仪上得到快速、可靠的结果。如果需要,反应液可用尿嘧啶DNA糖基化酶(UNG)预处理,以去除之前反应残留的PCR产物。

QuantiTect SYBR Green RT-PCR Kits配合QuantiTect Primer Assays可在基因表达分析中获得高度特异性的结果。基因组范围、生物信息学验证的引物对覆盖所有人类、小鼠、大鼠和其他各类生物基因。QuantiTect Primer Assays可在GeneGlobe网站上方便地订购。

Applications

QuantiTect SYBR Green RT-PCR Kit可用在各种real-time PCR仪上对cDNA目标序列进行基因表达分析,包括Applied Biosystems、Bio-Rad、Cepheid、Eppendorf、Roche和Agilent的仪器。在Rotor-Gene Q实时荧光定量PCR分析仪上使用时,我们推荐专门为快速PCR研发的Rotor-Gene SYBR Green RT-PCR Kit。

Feature
Specifications
Applications Real-time quantification of RNA targets
Reaction type Real-time and one-step RT-PCR
Real-time or endpoint Real-time
Sample/target type RNA
Single or multiplex Single
SYBR Green I or sequence-specific probes SYBR Green I
Thermal cycler All real-time cyclers (e.g. LC, RG, ABI)
With or without ROX With ROX

You are not authorized to download the resource

FAQs
33
FAQ ID -1056
View
FAQ ID -1083
View
FAQ ID -1084
View
FAQ ID -1085
View
FAQ ID -1086
View
FAQ ID -1141
View
FAQ ID -1143
View
FAQ ID -1146
View
FAQ ID -1438
View
FAQ ID -1441
View
FAQ ID -1446
View
FAQ ID -1447
View
FAQ ID -1451
View
FAQ ID -1714
View
FAQ ID -1982
View
FAQ ID -283
View
FAQ ID -2134
View
FAQ ID -2135
View
FAQ ID -2371
View
FAQ ID -2654
View
FAQ ID -2655
View
FAQ ID -2682
View
FAQ ID -312
View
FAQ ID -328
View
-20
View
FAQ ID -539
View
FAQ ID -551
View
FAQ ID -553
View
FAQ ID -564
View
FAQ ID -729
View
FAQ ID-751
View
FAQ ID -849
View
FAQ ID -850
View
Kit Handbooks
2
For genomewide, ready-to-use real-time RT-PCR assays using SYBR Green detection
Show details
For quantitative, real-time one-step RT-PCR using SYBR Green I
Show details