PCR – Withering Sydnrome cPCR Assay Limit of Detection Test

Ran cPCR, similar to that performed on 20130206, but with a lower set of dilutions, since the previous cPCR successfully detected down to 30 copies.

All samples run in duplicate.

Results:

Gel Loading (left to right):

Hyperladder V (Bioline)

30 copies

30 copies

15 copies

15 copies

10 copies

10 copies

5 copies

5 copies

3 copies

3 copies

NTC

A band is present in all samples, except the NTC. However, I’m frustrated by the appearance of the bands that these cPCRs are producing. I don’t like the smearing and looking at Nate’s poster outside of the lab, he has gel images with tight, very defined bands with no smearing. Will discuss with Lisa.

However, it’s interesting that the cPCR seems to detect all the way down to 3 copies! This suggests that the qPCR and cPCR don’t exhibit any difference in their limits of detection. I find this a bit odd, but will wait until I speak with Lisa to see what she says about Nate’s gel images.

UPDATE: Lisa indicated that the cPCRs are performed with different reagents/recipes than the qPCR! Will give those a shot to see if that improves the band quality.

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