i am going to make an invisible virus to become visible. all i have right now is its genomic sequence from a Genbank. this virus is non culturable as currently there is no suitable cell culture system to study its propagation. i don't have the viral stock/source, therefore there is no RNA extraction from any sample will be done.
is there any machine/method that i can use to synthesise RNA from the sequence?
please help because i am not familiar with PCR.. thank you ..
The main problem is that you don't have a template to transcribe. If you had the DNA you could perform an in vitro transcription. Since you only have the genomic sequence you could order the synthetic RNA (but that can be quite expensive) or have the essential genes that make the virus synthesized and cloned for HEK cells expression.
Hello!, yes, there are some methods to transcribe DNA to RNA. First you have to look for any plasmid that has T7, T4 or SP6 promoter (its more common for find T7 promoter). And look its MCS. Then design the Forward and Reverse primers with restriction enzymes to suit the right direction you need, then amplificate the genomic sequence you want with PCR reaction, then digest the amplicon, the acceptor vector and ligate. Afterwards you could do an in-vitro translation.
Regards,
Martin
- Badges
- Science topic