· ThermoSleuth软件职能：PCR验测设计工具 ? ? ? ?

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ThermoSleuth原名为ThermoBLAST，针对基因组几何扫描多个引物以查找全数杂交和扩增子。ThermoSleuth发现全数谬误杂交位点，扑灭PCR中的假阳性。

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解决引物异性

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ThermoSleuth凭据大型基因组数据库自动扫描寡核苷酸，以检测全数热力学不变的射中。与BLAST分歧，ThermoSleuth通过思考以下成分来捕获关键的谬误杂交射中：

• 适当的热力学评分，由于BLAST不会分辨GC对和AT对

• 显示全数假扩增子

• 思考3'的可扩大性和不变的错配

• ThermoBLAST凭据互补性而不是BLAST中的类似性进行评分

• ThermoBLAST允许溶液前提、盐、缓冲液、增长剂和其他尝试成分

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PCR假阳性问题

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PCR中假阳性的常见原因是由于引物杂交导致假扩增子的形成，而BLAST未检测到这种情况，由于它使用序列类似性而不是使用匹配和错配互补性的正确规定对射中进行谬误评分。此表，好多钻研人员无法获得现代基于基因组的分析设计所需的推算能力或存储空间。

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PCR假阳性的解决法子

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ThermoBLAST CE和ThermoBLAST通过使用BLAST的速度扫描人类基因组或微生物组等全基因组数据库，同时将适当的热力学模型利用于谬误杂交和交叉杂交了局，从而解决了引物和探针谬误杂交的问题。ThermoBLAST CE的云集成使全数拥有推算能力的钻研人员可能捕获50倍于BLAST的热力学不变和可扩大的射中。

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预造和可定造的基因组集中

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ThermoBLAST-CE蕴含一个重大的精选和预体式化序列数据库存储库，能够进一步分列成定造的序列播放列表。下面显示了一些受欢迎的播放列表？突Т丝棠芄辉诓幌薅饶诖妗⒋娲⒒蛲扑隳芰Φ那榭鱿鹿菇ㄋ璐笥椎暮蠖苁菘，这对于那些本地推算能力有限的钻研人员来说是一个巨大的疾苦起源。

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【英文介绍】

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Scans multiple primers against collections of genomes to find all hybridizations and amplicons.

ThermoBLAST finds all mishybridization sites, eliminates false positives in PCR.

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Solving Primer Specificity

ThermoBLAST automatically scans oligos against large genome databases to detect all thermodynamically stable hits. Different from BLAST, ThermoBLAST captures the important mishybridization hits by considering the following:

• Appropriate thermodynamic scoring, as BLAST doesn't differentiate a GC pair from an AT pair

• Displays all false amplicons

• Consideration for 3' extensibility and stable mismatches

• ThermoBLAST will score based on complementarity as opposed to similarity in BLAST

• ThermoBLAST allows for solution conditions, salt, buffers, additives and other experimental factors

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The problem of false positives in PCR

The most common cause of false positives in PCR is the formation of false amplicons due to primer mishybridization which is not detected by BLAST because it scores hits incorrectly using sequence similarity rather than using the proper rules for match and mismatch complementarity.? Additionally, many researchers do not have access to the computational capacity or storage required of modern genomic-based assay designs.

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The solution to false positives in PCR

ThermoBLAST CE and ThermoBLAST solves the problem of primer and probe mishybridization by scanning against whole genome databases such as the human genome or microbiome using the speed of BLAST while applying the proper thermodynamic model to mishybridization and?crosshybridization results. The cloud integration of ThermoBLAST CE enables all researchers with the computational power to capture 50X as many thermodynamically stable and extensible hits as BLAST.

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Pre-made and customizable genome collections

ThermoBLAST-CE contains a huge repository of curated and pre-formatted sequence databases that can further be arranged into a customized sequence playlist. Some of the most popular playlists are shown below. Customers now have the capability to build a background database as large as they need without restricting memory, storage or computational capacity which is a huge source of pain for those researchers that have limited local computational capacity.