Do not reuse old PCR for your control DNA : the control DNA controls many things, i.e. mix-up of primers, mix up of samples, gel loading errors, PCR quality, in addition to gel to gel variations.
Home page > MLVA Web service > Frequently asked questions > Do I need to do a PCR on the control strain each time a PCR for an "unknown" strain is setup? Or could I simply use leftover product from a previous PCR on the control strain to run on a gel with the products from the strains I am trying to type ?
Do I need to do a PCR on the control strain each time a PCR for an "unknown" strain is setup? Or could I simply use leftover product from a previous PCR on the control strain to run on a gel with the products from the strains I am trying to type ?
In the same section :
- I can get the PCR assay to work, but am having problems analyzing the bands to get accurate sizes from my gels. I wonder how to actually calculate an exact band size and the repeat number. I am using the size markers to make a standard curve based on distance migrated on the gel. This old fashioned method is working, but once I get a band size, I cannot figure out how to get a repeat copy number.
- May I run different loci on the same agarose gel ?
- I do not have the 16M reference strain. Is it possible to use an other control strain ?
- Is it better to use agarose or capillary electrophoresis for MLVA typing?
