I would check your spectrophotometry curves to make sure you don't have phenol contamination that is causing an overestimation of RNA quantity. Check for a shift in the peak toward 270 nm and a low 260/230 ratio.

Hii Anjan,

The RNA concentration of your sample ranging between 110-200 seems very low. I have 3 things to infer from your results;

1. The variation is can occur due to undissolved remains in the sample for that keep the RNA sample in an incubator at 55'C for 5 mins.

2. However, for RNA content from next time onwards take more salvia sample for RNA isolation, also reduce the final MQ water volume to dissolve it.

3. Prepare cDNA with 500 to1000ng of RNA content, thereby you may get better amplification of your desirable gene in it. For present sample you can try with double or more volume of cDNA in PCR.

Best wishes

Hello Anjan Behera

In addition to what John Hardy Lockhart and Dinesh Raj Pant mentioned, I would suggest you run your RNA samples (500-1000ng each) in a 1% bleach gel or formaldehyde agarose electrophoresis. That will help you visualize if you have intact RNA. Only if you see intact RNA then your amplification of GAPDH and ACTB will make sense.

Hope this helps.

Best!

you can also perform a RIN ( RNA integrity number) by DHPLC ( Agilent)

because you can detect RNA but it's very fragmented.

If it's the case you can use a RNA kit( Thermo) to repair some gaps, if the fragmentation, is not dramatic. But if it was all this case, you fast collect a lot of

saliva 10 or 20 ml and immediately freeze it in trizol

Hi

The RNA seems fragmented, so the possibility of getting PCR amplification of genes is low. See if some published literature offer tips regarding RNA isolation from saliva samples...if you get good integrity of RNA then only proceed further for anlysis