During the reverse transcription step, you may use a single primer to generate cDNA. However, to amplify for use in subsequent cloning or sequencing steps you should perform PCR on the cDNA generated. 

Ya i get it..I was worried only about the reverse transcription step. oligo dT is not giving me my complete transcript, so I was wondering if I could use gene specific primer.

Yeah, I usually start with the oligo dT when amplifying cDNA and then follow up with specific primers for PCR. However, if this doesn't work (which happens on occasion with larger amplifications) then I use a specific primer for the initial cDNA reverse transcription step. Just make sure that you have plenty of dNTPs available and that your enzyme is active and it should work fine. Good luck.

i believe random primers should be used instead of oligo dT for your RT, when you want to retrotranscribe long transcripts.

Hello, I have cloned a transcript bigger than 8300bp making the Reverse Transcription with specific primier. However I think that you can use specifics primers or random primers or oligo dT for the RT. I think that is more important the quality of the total RNA (or mRNA), the kind of transcriptase, the Tº and the times that you are using for the RT.

Did you check your RNA quality by agarose gel?

Which transcriptase are you using? I recommend you SupreScript (I used this enzime), or another enzime without RNase H activity (or reduced RNase H activity), to prevent RNA degradation by the transcriptase. Or at least an enzime that works properly a high temperature 50 or 55ºC for example, to avoid the secondary RNAs structure.

If you use random or oligo dT primers, start the transcription 10 min at 37ºC, then 10 min at 42ºC and finally 50 min at 55ºC.

Good luck!!!!

Christian

thank you Mr. Martin Moya. i do check the quality on a gel before proceeding. I use revert aid reverse transcriptase.Shall take all your suggestions into consideration before I proceed.