PCR; Perhaps someone can explain? - PCR; Primer (May/25/2005 )
Hi, I'm a student at a high school in Holland. I recently got an assignment from school to isolate IL-6 mRNA from T24 celline. (human bladder carcinoma cells) and then make cDNA from the IL-6 RNA.
Now my approach would be to use a kit (Oligotex from QIAGEN) to purify the RNA from the cells. Then i want to use the iScript cDNA Synthesis kit (BIO-RAD) to make the cDNA.
But i think need to isolate the IL-6 mRNA from the other mRNA first by using PCR.
For that i need a primer for IL-6.
How do i find the primer for this purpose???
I have never done a PCR before although i'm familiar with the theorie.
Thx,
Junior
P.S. Feel free to make any suggestions in how to tackle this assignment.
Now my approach would be to use a kit (Oligotex from QIAGEN) to purify the RNA from the cells. Then i want to use the iScript cDNA Synthesis kit (BIO-RAD) to make the cDNA.
But i think need to isolate the IL-6 mRNA from the other mRNA first by using PCR.
For that i need a primer for IL-6.
How do i find the primer for this purpose???
I have never done a PCR before although i'm familiar with the theorie.
Thx,
Junior
P.S. Feel free to make any suggestions in how to tackle this assignment.
You may search Pubmed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi) for earlier publications on IL6 that might have primer info in them. Alternatively you can search GenBank (http://www.ncbi.nlm.nih.gov/Genbank/) for sequences of human IL6 and use any of the freeware primer designing programmes (this website also has quite a few of them) and get your specific primers designed.
All the best.
Sree
How to design rRT-PCR primer? Here is a tutorial http://www.protocol-online.org/forums/inde...?showtopic=2706
Thanks i'll look into it right away.
Junior