problems extracting RNA from DCs

[ateijeiras]

I'm extracting RNA from cultured differenciated human Dendritc cells using TRI zol and i'm having very low amounts of purified RNA. I don't have more than a million of cells. do anyone know what could be the problem? Have this cells litle amount of mRNA? i would really appreciate any suggestion. I´m new on this
thank you
Álvaro

[El Gú]

Dear Alvaro, i work with macrophages in culture bottles or nuncl plaques. I do some like that: i wash the culture with PBS and a put directly the trizol. I colet the trizol and follows the rna extraction protocols. On isopropanol phase i used LPA (linear polyacrilamide) (+- 5ul, per 700 ml of trizol), this reagent drop small nucleic acid like dna less than 20pb. Using this reagent i obtain much quality of my rna amounts. I hope that this information help you.

Álvaro one favor, can you send me the isolation of CDs of any tissue. I like work with gingival tissues.

Thank alot.

Gustavo.

[El Gú]

Dear Alvaro, i work with macrophages in culture bottles or nuncl plaques. I do some like that: i wash the culture with PBS and a put directly the trizol. I colet the trizol and follows the rna extraction protocols. On isopropanol phase i used LPA (linear polyacrilamide) (+- 5ul, per 700 ml of trizol), this reagent drop small nucleic acid like dna less than 20pb. Using this reagent i obtain much quality of my rna amounts. I hope that this information help you.

Álvaro one favor, can you send me the isolation of CDs of any tissue. I like work with gingival tissues.

Thank alot.

Gustavo.