qPCR – RLOv DNA Helicase with Black Abalone DNA with Varying Levels of RLO/RLOv Repeated

Repeated the qPCR from 20151120 because the last attempt I did not have samples that had been shown to be clean of RLO via qPCR; had used histology scoring to identify RLO-negative samples. Additionally, many of the samples that produced amplification were out of range of the standard curve (most came up too early) and require dilution to be properly assessed.

RLO/RLOv 0 RLO/RLOv 1 RLO/RLOv 2
08:4-3 06:5-35 06:5-31
08:4-4 06:6-32 06:5-32B
08:4-5 06:6-39 06:6-46
08:4-6 06:6-42 06:6-49
08:4-7 06:6-44 08:3-05
08:4-8 06:6-52 08:3-07
08:4-9 06:6-54 08:3-15
08:4-10 06:50-08 08:3-16
08:4-11 06:50-10
08:4-12 07:12-18

Made 1:1000 dilutions of all of the RLOv 1 and RLOv 2 samples.

Master mix calcs are here (Google Sheet): 20160106 – qPCR RLOv black abs

Ran qPCR using the RLOv DNA helicase standard curve from 20151106.

All samples were run in duplicate.

Plate layout, cycling params, etc., can be found in the qPCR Report (see Results below).

Results:
qPCR Report (PDF): Sam_2016-01-06 16-01-36_CC009827.pdf
qPCR Data File (CFX96): Sam_2016-01-06 16-01-36_CC009827.pcrd

Summary:

Standard curve looks solid.

RLO-negative samples are all negative for RLOv DNA helicase.

1:1000 dilutions of RLOv 1 and RLOv 2 all amplified within the range of the standard curve.

 

RLO Negative Amplification Plots

 

RLO 1 Amplification Plots

 

RLO 2 Amplification Plots

2 thoughts on “qPCR – RLOv DNA Helicase with Black Abalone DNA with Varying Levels of RLO/RLOv Repeated

  1. Pingback: Data Aggregation – WS RLO and RLOv DNA Helicase qPCR and WS RLO Infection Intensities | Sam's Notebook

  2. Pingback: qPCR – Phage Portal Primer Specificty Check | Sam's Notebook

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