Daniel:Notebook/PosSequencing/2017-4-28

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Emulsion PCR (Started Yesterday)[edit]

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Images and Analysis[edit]

Field 4[edit]

Field 4 had the best results and is worth a summary. There are 5 fluorescent spots. Not counting Cy3, which shouldn't be there and has extra signal that is not present in the other channels. Two of the molecules are too large to be a single bead, which should be ~1 um. However, 3 of the signals are that size. Two have A488 signal and one has Cy5 signal.

Other Fields[edit]

Conclusion[edit]

In conclusion I don't think the experiment had much conclusive results. There do appear to be several beads that monoclonal and are the right size. There is also significant Cy3 signal that shouldn't be there and Cy5 seems weak. Only 1 Cy5 bead was suggested in field 4, whereas 2 A488 beads were in a 4 and possibly another 2 in field 2. Recommend repeating without the emulsions and with only single fluorophores (1 each) and a mixed channel to show what can happen when the beads are mixed/unmixed. Basically, a bunch of positive controls for comparison.