Can you send some more details of the protocol you followed for analysis of your gene and a picture of the 'curve' you have generated in qRT-PCR...?

Alright. I extracted total RNA using a Promega kit. After that I measured RNA concentration in nanovue. Next, I've transcribed 400ug do RNA into cDNA using TaqMan reagents. The cDNA obtained was diluted five times. Than I take the control and I made a serial dilution - 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, and the numbers 500, 1000, 2000, 4000, 8000, 16000 was atributed to each dilution respectively. 2uL of each point was pipetted with 2.4uL of water, 5.0uL of SYBR, 0.3uL of each primer 10uM. In a total volume of 10uL. The themarl profile was: 50°C for 2min; 95°C for 10min; and 40 cycles of 95°C for 15 sec and 60°C for 1 min. I'll put the curves of Actin and two different primers of BDNF.

BDNF first pair of primers

BDNF second pair of primers

Hi Vicente......Thanks for uploading the images....After going throught the description and the images of the qRT-PCR ....I will give my views tommorrow in an attachment as it may be a long message to write here....furthermore I also have few more queries....which also I'll inquire in the attachment

Thanks for sendingthe images...I am sure something will be sorted out...

bye

Hi Vicente

see my comments and suggestions in the attachment......do feel free to contact if you need some clarification...bye

Just doing some corrections, I've used 400ng of total RNA.