Sam:LabNotes/Microbione/2009-2-5: Difference between revisions

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>Sam Chiang
>Sam Chiang
Line 21: Line 21:


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


==Suggestion==
==Suggestion==

Revision as of 19:46, 5 February 2009

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