|
|
|
How do I avoid false positive and false negative
|
|
The false negative results usually come from bad DNA
quality or failed PCR. Internal control is the best solution to prevent
this problem. In IQ2000TM system, internal control (house-keeping gene) is
included and users can use the internal control to monitor the DNA quality and
PCR procedure easily.
The false positive results usually come from
contamination of the final PCR products or cross contamination during
extraction and reagent-adding step. To avoid contamination from the PCR
products, the best way is to isolate the gel electrophoresis room far away from
the sample preparation and reagent preparation room. About the cross
contamination problem, a good training is the best way to prevent it.
Back to "Q&A"
|
|
How do I know the sensitivity is appropriate? Can I adjust the sensitivity?
|
|
You can monitor
system's sensitivity by the quantified positive standard included in the
kit. In IQ2000TM WSSV system, you should dilute the standard to 10 copies/ul
by yeast tRNA. Then you can monitor the sensitivity by the 10-copy
standard. If the 10-copy standard resulted 3 bands (910, 550, and 296 bp),
your system is over sensitivity. You may reduce 2 to 3 cycles from the
first reaction. On the other hand, if the 10-copy standard did not result
any band, you may increase 2 to 3 cycles on the first reaction to increase the
sensitivity.
Back to "Q&A"
|
|
Must I see the hose-keeping gene's product from the negative samples?
|
|
It depends on what
kind of sample you want to test. Because the house-keeping gene was
designed according to the shrimp genomic DNA, only the shrimp samples will show
the right products. The negative shrimp samples must show the
house-keeping gene; otherwise, you have to check the DNA quality to avoid false
negative results. For other crustaceous animals, they might show the house
keeping gene but the product size could be different from the 848 bp in some
species. If the samples are pond dirt, pond water, or feed, you will not
get the house-keeping gene.
Back to "Q&A"
|
【Product】【Virus Info】【News】【Service】【Q&A】【About us】【Distributors】
|