Good afternoon all. I'm a PhD student from India currently working on scrub typhus disease investigation. I extracted DNA from blood samples using Qiagen blood extraction kit and screened by using conventional PCR. The samples which were positive were stored at -20 degree centigrade. After a span of 2 to 3 months, I again screened them using same primers and reaction conditions but they didn't appear positive except one sample which showed very faint band. what could be the possible reason for this? Is this because the scrub typhus genomic DNA is unstable? or because of wrong chemicals used in processing samples or because of wrong storage conditions? Kindly help me with this research problem if anyone can shade light on this aspect.
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