MeltDoctor™ HRM 预混液
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Applied Biosystems™

MeltDoctor™ HRM 预混液

Applied Biosystems™ MeltDoctor™ HRM 预混液含有实现 HEM 卓越性能的所有组分(模板和引物除外),广泛适用于各种基因组靶点。高分辨率熔解了解更多信息
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货号数量
44154405 mL
440953550 mL
441545010 x 5 mL
44154525 x 5 mL
货号 4415440
价格(CNY)
5,196.00
Each
添加至购物车
数量:
5 mL
价格(CNY)
5,196.00
Each
添加至购物车
Applied Biosystems™ MeltDoctor™ HRM 预混液含有实现 HEM 卓越性能的所有组分(模板和引物除外),广泛适用于各种基因组靶点。高分辨率熔解(High Resolution Melting,以下简称 HRM)分析是一种后 PCR 分析方法,可用于鉴别核酸序列的基因变异。与其它市售预混液不同,MeltDoctor™ HRM 预混液在使用前无需混匀,专为 HRM 应用而开发和优化。
•背景荧光低
•在双链 DNA 存在条件下具有高亮度
• 因染料结合引起的 DNA 熔解温度变化极小
• 具有热稳定性,能耐受 PCR 循环条件
• 不抑制 DNA 聚合酶活性,PCR 效率高
• dNTP 混合物中包含 dUTP,它利用尿嘧啶 DNA 糖基化酶 (UDG) 在后续 PCR 反应中降解扩增片段,可大大减少残留污染

进行突变筛查
HRM 分析可用于筛查靶基因突变,在测序分析前鉴别变异样本。HRM 作为一种突变筛查技术,与诸如 DHPLC 和 DGGE 等常规方法相比具有明显的优势。HRM 用于突变筛查的优势包括:
• 试剂消耗量低,几乎没有浪费:HRM 仅需一个 20 µL PCR 反应即可分析单个样本,无需使用 HPLC 溶剂或 DGGE 凝胶
• 流程简单快速:PCR 扩增完成后无需使用其它仪器。只需在 PCR 图谱分析后添加高分辨率熔解曲线步骤,即可立即分析。
• 快速优化:不同于 DHPLC,无需热优化
• 样本用量低:经 HRM 分析后的 PCR 产物可直接用于 Sanger 测序反应

高质量组分
• 精确的镁盐及其它缓冲组分配方,可获得良好 HRM 结果。
•AmpliTaq Gold™ 360 DNA 聚合酶是一种高度纯化的 DNA 聚合酶,具有热启动性能,可大大减少非特异性产物形成并可在室温下反应
• MeltDoctor™ HRM 染料是一种稳定的荧光 SYTO™ 9 双链核酸染料,由 Molecular Probes 开发。
仅供科研使用。不可用于诊断程序。
规格
适用于(设备)7500 Fast 系统、7900HT Fast 系统、QuantStudio™ 12k Flex、QuantStudio™ 3、QuantStudio™ 5、QuantStudio™ 6 Flex、QuantStudio™ 7、StepOne™、StepOnePlus™、ViiA™ 7 系统
基因分型靶标SNP(未知或多种),表观遗传学靶标
反应次数500 次反应
聚合酶AmpliTaq Gold 360 DNA Polymerase
产品线MeltDoctor™
产品类型HRM Master Mix (2X)
数量5 mL
样品类型DNA(基因组)、cDNA
运输条件湿冰
技术高分辨率熔解 (HRM) 分析,突变筛查
容量5 mL
检测方法HRM
适用于(应用)基因分型
形式液体
高 GC PCR 扩增效果
标签或染料HRM(高分辨率熔解)染料, SYTO 9
PCR 方法qPCR
反应速度快速
Unit SizeEach
内容与储存
MeltDoctor™ HRM 预混液的组分为 2X 浓度,便于使用,包括:AmpliTaq Gold™ 360 DNA 聚合酶,MeltDoctor*trade;HRM 染料,一种含 dUTP、镁盐和其他缓冲液组分的 dNTP 混合物,配制精确,能提供良好的 HRM 结果。

储存于 -15 °C 至 -25 °C 下

保证的最短有效期为60天(打印于产品和检验报告上的确切有效期)。

常见问题解答 (FAQ)

The HRM software will not accept my calibration file. What is wrong?

Check that there are no outliers on the plate. You cannot omit any wells on the HRM calibration plate.

I have a 7900HT Fast Real-Time PCR System, and cannot open the data file in HRM software. What could be causing this?

There are a few possibilities. First, make sure the HRM Software version is v2.0.1, and the 7900HT Fast Real-Time PCR System software version is v2.3 or above. Second, check that the run method used was as recommended in the HRM protocol; make sure the ramp rate for the dissociation stage is 1%. Then try to open the calibration file from the HRM Software; if it does not open, the calibration file is defective. The defects could be due to a bad calibration plate or instrument uniformity issue.

My 7500 Fast Real-Time PCR System has been upgraded to v2.0.6, so why can't I analyze my files with HRM Software v2.0.1?

For the 7500 Fast Real-Time PCR System, if the software version is below v2.0.4, the compatible HRM Software is v2.0.1. If the 7500 Fast Real-Time PCR System has been upgraded to software version 2.0.4 or above, then you will need to get HRM Software version 3.0.1.

How can I get positive controls for the different genotypes for my HRM experiment?

You can order a synthetic positive control plasmid using our GeneArt Gene Synthesis services. The final product is sequence verified and provided as 5 µg of lyophilized plasmid. Alternatively, you may be able to find a positive control sample from Coriell.

How do I perform the HRM calibration on my real-time PCR instrument?

The HRM calibration is a 3-step procedure. In some instrument software, the calibration process has been streamlined to a single-step procedure (applies to the ViiA7 Real-Time PCR System and QuantStudio Real-Time PCR Systems).

For the HRM calibration, the first step is a PCR reaction to generate the HRM dye plate. The reaction plate contains a DNA template, a primer pair, and the PCR mix with HRM dye. The second step is dye calibration using the PCR plate from step 1. The last step is a melt curve run, again using the same plate. For details, please refer to the HRM guides for each instrument:

- 7500 Fast Real-Time PCR System (with software version 1.4 or 2.0.4 and below) (https://tools.thermofisher.com/content/sfs/manuals/cms_050347.pdf)
- 7500 Fast Real-Time PCR System (with software version 2.0.5 and above) (https://tools.thermofisher.com/content/sfs/manuals/4457847.pdf)
- 7900HT Real-Time PCR System (https://tools.thermofisher.com/content/sfs/manuals/cms_050347.pdf)
- StepOne and StepOnePlus Real-Time PCR Systems (https://tools.thermofisher.com/content/sfs/manuals/4457847.pdf)
- ViiA 7 Real-Time PCR System (https://tools.thermofisher.com/content/sfs/manuals/4443531.pdf)
- QuantStudio 6 and 7 Real-Time PCR Systems (https://tools.thermofisher.com/content/sfs/manuals/4489823.pdf)
- QuantStudio 6 and 7 Pro Real-Time PCR Systems (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0018981_HighResolutionMelt_DA_SW_UG.pdf)
- QuantStudio 3 and 5 Real Time PCR Systems (https://tools.thermofisher.com/content/sfs/manuals/MAN0014394_HighResMeltExperiment_GSG.pdf)
- QuantStudio 12K Flex Real-Time PCR System (https://tools.thermofisher.com/content/sfs/manuals/4470692A.pdf)

Find additional tips, troubleshooting help, and resources within our High Resolution Melt Support Center.