Biotechnology Forums

Hi,
I've done gene transformation to rice.To confirm that new gene has been transferred to rice I've already done genomic DNA PCR and the result showed the existence of the new gene but My supervisor asked me to do RT-PCR (Reverse transcription) too. Could someone please let me know what are RT-PCR preferences to genomic DNA? When I got the desire result by total genomic PCR, what's the point to do RT-PCR?
Thanks.

(01-30-2012, 02:00 AM)charisma Wrote: [ -> ]Hi,
I've done gene transformation to rice.To confirm that new gene has been transferred to rice I've already done genomic DNA PCR and the result showed the existence of the new gene but My supervisor asked me to do RT-PCR (Reverse transcription) too. Could someone please let me know what are RT-PCR preferences to genomic DNA? When I got the desire result by total genomic PCR, what's the point to do RT-PCR?
Thanks.

Maybe he want to study how the gene expression works on the mRNA level. This method allows to copy sequence which is only included in mature mRNA (exons).

He/she wants you to study the expression level of that gene of interest that has been transformed into your rice genome. Also, the result you've obtained will help you understand the expression patterns of the randomly selected plantlets. You don't want transgene silencing to occur in your transformed rice, no?

Hope it helps! =)

Because it works and give better results