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AlleleID 物种识别和分类鉴别的综合软件
的实验设计
AlleleID是一款旨在解决、病原体检测或物种的综合性桌面工具。以CulsSTW多序列比对为核心,AlleleID可以设计用于物种识别/交叉物种的基因芯片探针以及用于实时PCR的SYBR Green, TaqMan MGB, TaqMan探针, Molecular Beacons以及实时PCR引物。AlleleID还提供了支持可变剪切片段检测的微阵列实验设计。
功能特征
物种试验
基因芯片的基因拼接/选择性剪接
外显子连接引物设计
实时PCR引物设计
TaqMan探针设计
SyBR绿色底漆设计
分子信标设计
FRET探针设计
微阵列探针设计
精密化验设计
数据库支持
输入输出格式
ross Species/Taxa Specific Assays
Species Identification Assays
Gene Splicing/Alternative Splicing using Microarrays
Exon Junction Primer Design
Real Time PCR Primer Design
Real Time PCR
Introduction to Real Time PCR
As the name suggests, real time PCR is a technique used to monitor the progress of a PCR reaction in real time. At the same time, a relatively small amount of PCR product (DNA, cDNA or RNA) can be quantified. Real Time PCR is based on the detection of the fluorescence produced by a reporter molecule which increases, as the reaction proceeds. This occurs due to the accumulation of the PCR product with each cycle of amplification. These fluorescent reporter molecules include dyes that bind to the double-stranded DNA (i.e. SYBR® Green ) or sequence specific probes (i.e. Molecular Beacons or TaqMan® Probes). Real time PCR facilitates the monitoring of the reaction as it progresses. One can start with minimal amounts of nucleic acid and quantify the end product accurately. Moreover, there is no need for the post PCR processing which saves the resources and the time. These advantages of the fluorescence based real time PCR technique have completely revolutionized the approach to PCR-based quantification of DNA and RNA. Real time PCR assays are now easy to perform, have high sensitivity, more specificity, and provide scope for automation. Real time PCR is also referred to as real time RT PCR which has the additional cycle of reverse transcription that leads to formation of a DNA molecule from a RNA molecule. This is done because RNA is less stable as compared to DNA
Limitations of End-point PCR
In a PCR reaction as the reaction progresses, the reagents are being consumed as a result of amplification. Now the PCR product is no longer being doubled at each cycle due to this reagent constraint. This depletion will occur at different rates for each replicate. Thus, the samples begin to diverge in their quantities. This diminished amplification is the linear phase of the reaction. The plateau for each tube will differ due to the different reaction kinetics for each sample. It is in this phase where traditional PCR takes its measurement, also known as the end-point. This End-Point Detection has some problems such as low resolution, poor precision, low sensitivity and the need for post PCR processing. Also, the results of this detection are not expressed in numbers and there is no scope for automation.
AlleleID® is a comprehensive desktop tool designed to address the challenges of bacterial identification, pathogen detection or species identification.
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