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I am Malek, Master student in UPM, Malaysia. currently, I am doing my research about Bacterial panicle blight (BPB) caused by Burkholderia glumae bacteria that associated with rice in Malaysia. After do isolation and biochemical test, I ran PCR by using universal primers (16s). Unfortunately, after Blast the sequences result, there is no Burkholderia glumae detected, but mostly Enterobacter. My supervisor said, try to use specific primers of Burkholderia glumae and do gradient pcr in order to check which annealing temperature is suitable to detect Burkholderia glumae when annealing process occurred. But, after i ran, there is no any bands appeared. I've checked and troubleshoot about concentration and pcr program and so on, but still cannot figure out the problem. Hope you can help me by suggesting other ways.

Thank you. Heart
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