Currently I am working with qPCR quantification specific for Lactobacillus bacteria. I found in some cases, the sample showing multiple peaks of melting curves while others looks perfect, so do the amplification plot duplication. Any one have an opinion about this issues? Does it caused by sample DNA quality, the omount of the bacteria detected, or primers problem? I am condisering that this condition reflecting the low abundance of the specific bacteria inside its sample.
Currently I am working with qPCR quantification specific for Lactobacillus bacteria. I found in some cases, the sample showing multiple peaks of melting curves while others looks perfect, so do the amplification plot duplication. Any one have an opinion about this issues? Does it caused by sample DNA quality, the omount of the bacteria detected, or primers problem? I am condisering that this condition reflecting the low abundance of the specific bacteria inside its sample.
Biomedical Instrumentation Systems
1st Edition
ISBN:9781133478294
Author:Chatterjee
Publisher:Chatterjee
Chapter15: Instrumentation In Medical Imaging
Section: Chapter Questions
Problem 10P
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Question
Currently I am working with qPCR quantification specific for Lactobacillus bacteria. I found in some cases, the sample showing multiple peaks of melting curves while others looks perfect, so do the amplification plot duplication. Any one have an opinion about this issues? Does it caused by sample DNA quality, the omount of the bacteria detected, or primers problem? I am condisering that this condition reflecting the low abundance of the specific bacteria inside its sample.
Transcribed Image Text:Amplification Plot
1000
100
10 10.0
0.1
0.01
0.001
0.0001
0.00001
0.000001
2
4
10
12
14
16
18
20
22
24
26
28
30
32
34
36
38
40
Cycle
Legend
A
D
E
G.
ARn
Transcribed Image Text:Melt Curve
760000.0
660000.0
580000.0
460000.0
360000.0
260000.0
160000.0
60000.0
-40000.0
85.0
70.0
75.0
80.0
85.0
90.0
95.0
Tm: 82.91
Temperature ("C)
Legend
В
D
E IF
Derivative Reporter (-R)
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