Hi Dear
when i going to perform PCR , i get the band for some samples while not for the rest of the sample. the gene is housekeeping gene so it must be present in all samples
Hi Dr. Hussein,
Same question that always asked, the disappearance of bands of housekeeping genes in some samples, and one possible answer in my mind is the DNA in range of this gene in some samples got breaked
Hello Dr. Hussein Oleiwi Muttaleb Al-Dahmoshi
Primarily, you can dilute your template DNA serially like 1:10 and 1:50 and 1:100, set up PCR. analyse the data and prove the presence of inhibitors if any.
Secondly, elimination of inhibitors depends on the type of sample. If it is sputum sample from where you isolated the template, you could use NALC for decontamination.
Dear Dr. Hussein Oleiwi Muttaleb Al-Dahmoshi,
it is important to use DNA extraction kit, that is designed to your sample type. Make sure it is your case, and make sure the extraction in all samples is done by excatly the same protocol. Ideally get measurement of DNA concentration in youe extractions.
The PCR inhibition can be reduced by dilution of the sample as already suggested.
Bovine serum albumin is used to reduce inhibition in qPCR, (e.g. Article BSA reduces inhibition in a TaqMan (R) assay for the detecti...
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