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problems extracting RNA from DCs

problems extracting RNA from DCs - RNA Techniques Forum

problems extracting RNA from DCs - Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols.


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  #1  
Old 01-22-2009, 05:56 PM
Pipette Filler
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Default problems extracting RNA from DCs



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
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Old 04-01-2009, 01:27 AM
Pipette Filler
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Default Re: problems extracting RNA from DCs

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.
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Old 04-01-2009, 01:27 AM
Pipette Filler
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Default Re: problems extracting RNA from DCs

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.
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