These two genes are on the same bacteria genome DNA. When I did a multiplex PCR, for some samples, both of them will show up, but for some other samples,gene 1 will show when the template is around 500ng, while gene 2 will only show when the template is diluted to 50ng. However, when the DNA diluted to 50ng, gene 1 wont show up anymore.
I assume gene 1 is low in abundance, so it needs higher DNA concentration, while gene 2 is inhibited by something, then it requires dilution. Is it possible? if so how come gene 1 is not inhibited?
why the abundance of two genes is so random among different samples?
Hi Liang,
Several valid suggestions are given above by Briony. They explain the type of interactions happening between the primer and template and the factors that can affect them. Such non-consistent amplifications are not solely related to gene copy number, instead among many factors the quality of template preparation will also play a major role...
bye
Thanks a lot for the answers Briony and Ajay.
I am starting with improving template quality by trying different isolating methods.
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