Hello, all over the internet there are explanations that leave out important parts of how things are done (or worse: that are even wrong?). Does anyone have a good source? I am especially interested in this:
How do you make sure, that you assign only one molecule to one bead? If say the adaptor of molecule A hybridises with the probe on the bead, what would keep another molecule B from hybridising with another probe on that bead?
Does the A adaptor anneal to the 5' end and the B adaptor to the 3' end?
Is the probe on the bead complementary to the A or the B adaptor?
Does the bead have only A probes or only B probes annealed to it or both?
How in detail does the amplification on the bead work? I see pictured explanations where on one picture 1 molecule attaches to the bead and on the next picture suddenly there's many of them. And the explanation states "It's PCR". But that's not enough.
I have played through so many scenarios and every time I get stuck somewhere thinking "But that cannot work because of bla". So I would really appreciate a source of a detailed explanation.