Sam:LabNotes/Microbione/2009-2-5

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Revision as of 19:46, 5 February 2009 by >Sam Chiang (→‎Discussion)
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Follow up: Testing human 18S and Bac (E. coli) 16S primers using realtime PCR

Exp. Design

Template         gDNA      H2O
                --------  ---
Primer: 18S-211  #1   #2   #3    (#1, #2 - replicates)
        18S-306  #4   #5   #6    (#4, #5 - replicates)


Results

FIG.1 Realtime amplification plot (no log in flourescent)

File:Sam020409.jpg


FIG.2 Realtime amplification plot (with log in flourescent)

File:Sam020409-log.jpg

Discussion

  1. The amplification curves are not distinguishable (Sample vs. Blank) within 20 clycles.
    1. The amplifications cycless were not enough?
    2. The amplifications are not working?
      1. Primers' specificity? binding affinity?
      2. Concentration of templates?

Suggestion