After a run, you get a quality report of the loading of a particular cell, that is broken down into: 2, 1 and 0.
My question pertains to what constitutes a 2?
My understanding of the SMARTbell adapts and ligation biochemistry is such that every closed ss-circle will be flanked by two SMARTbells with the same adapter = that there are 2 priming sites per closed circle.
How is priming and amplification from BOTH priming sites prevented?
Does a '2' correspond to two separate circles loading into the ZWM (overloading) or two different sequences arising from a ZMW, possibly from independent priming off the redundant sites in the same circle (concentration of primer may be off)?
Thanks.
My question pertains to what constitutes a 2?
My understanding of the SMARTbell adapts and ligation biochemistry is such that every closed ss-circle will be flanked by two SMARTbells with the same adapter = that there are 2 priming sites per closed circle.
How is priming and amplification from BOTH priming sites prevented?
Does a '2' correspond to two separate circles loading into the ZWM (overloading) or two different sequences arising from a ZMW, possibly from independent priming off the redundant sites in the same circle (concentration of primer may be off)?
Thanks.
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