PCR, qPCR and mPCR
No one can imagine biotech without PCR. There are many variations of the basic PCR technology, just visit the Wikipedia link, to find more than 20 different PCR based methods. qPCR or real time PCR, is one of the most popular technologies today. It is popular because it has several desired features. It is sensitive, [...]
Read morePPI helps you design better PCR primers
PPI stands for polymerase preference index (patent pending). It is an algorithm we developed to help select the best performing PCR primers based on polymerase preference. Most of the primer design softwares evaluate the melting temperature (TM) as the most important parameter. But TM, at its best, could only tell us if a primer would [...]
Read moreiCubate 2.0, an open platform for assay developers
iCubate 2.0 is an idea we borrowed from the IT industry where the Web 2.0 business model has already transformed our lives. Here, the “2.0” means “user generated content and an online store”. So, iCubate 2.0 is an open platform that allow users to develop and market multiplex PCR (mPCR) assays. We call this kind [...]
Read moreWhy we care about the design of the instrument?
Walking into a clinical molecular diagnostic lab, you will see many boring, gray, ugly shaped machines. It seams nobody is willing to make that little extra effort to design a better looking instrument. The plastic shells are almost always an after thought, simply to cover up the guts. With iCubate, we paid attention to aesthetic [...]
Read moreiCubate cassette improves sensitivity with a little trick
We really treat the iCubate cassette like slaves: we tell them do many tedious and “labor intensive” things without giving them a break, and they are not allowed to make any mistakes. One of the things we ask the cassette ( or more precisely, the pipette in the cassette) to do lately is to add [...]
Read moreClosed iCubate cassette may give you a new office
Typically, to set up a molecular lab, you need at least three isolated rooms: one for sample prep, one for setting up the PCR reactions, and one for detection. You need these rooms because of the concern about amplicon contamination that leads to false positive results. That concern is warranted, because like earthquakes in California, [...]
Read moreWhat it takes to automate an assay?
The answer: (1) start with a simple assay that really works; (2) finish it by a great engineer team. There were several multiplex attempts out there, but non become successful commercial product. The reason: the assay they try to automate usually has too many enzymatic steps, and includes too many procedures such as post-PCR clean [...]
Read moreMultiplexed assays have greater clinical sensitivity
There are two kinds of sensitivities: analytical sensitivity, and clinical sensitivity. Analytical sensitivity is all about copy numbers: How many copies would your test need to give a positive result? The more sensitive an assay is, the less copy it will require to yield a positive result. Clinical sensitivity is all about the patient: when [...]
Read moreReinventing PCR: Who?
Why us? why we were the lucky ones that developed the new multiplex PCR strategies? The Polymerase Chain Reaction was invented more than 20 years ago, the inventor, Kary Mullis, won Nobel Price in 1993. I believe that the reason why so many people failed to make multiplexing work is because we all trusted the [...]
Read moreReinventing PCR: How?
To solve the three problems mentioned before, we came up with this multiplex PCR strategy called tem-PCR (for target enriched multiplex PCR). We use at least two pairs of nested primers for each amplification target, so if there is 10 targets we want to amplify together (multiplex), there will be total of 40 primers (4×10). [...]
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