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Carmen Sandoval
Carmen Sandoval
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I am currently conducting an experiment which involves FACS-sorting a specific population of cells using an antibody of interest.

In order to validate the type of cells I have collected using this marker, I want to perform qPCR on their extracted RNA, using certain known cell-type marker primers.

My problem is that in each sorting session, I am only able to collect anywhere from 5,000 - 10,000 cells. Do you know if it is possible to perform RT-qPCR on such a small sample size?

I have collected the cells in 1mL of Trizol.

Would I maybe have to amplify the cDNA after reverse transcription in a general manner ( using random primers)?

Thanks for your help.

I am currently conducting an experiment which involves FACS-sorting a specific population of cells using an antibody of interest.

In order to validate the type of cells I have collected using this marker, I want to perform qPCR on their extracted RNA, using certain known cell-type marker primers.

My problem is that in each sorting session, I am only able to collect anywhere from 5,000 - 10,000 cells. Do you know if it is possible to perform RT-qPCR on such a small sample size?

I have collected the cells in 1mL of Trizol.

Thanks for your help.

I am currently conducting an experiment which involves FACS-sorting a specific population of cells using an antibody of interest.

In order to validate the type of cells I have collected using this marker, I want to perform qPCR on their extracted RNA, using certain known cell-type marker primers.

My problem is that in each sorting session, I am only able to collect anywhere from 5,000 - 10,000 cells. Do you know if it is possible to perform RT-qPCR on such a small sample size?

I have collected the cells in 1mL of Trizol.

Would I maybe have to amplify the cDNA after reverse transcription in a general manner ( using random primers)?

Thanks for your help.

Source Link
Carmen Sandoval
Carmen Sandoval
  • 321
  • 4
  • 10

How to Extract RNA and perform RT-qPCR from very few cells ( ~5,000)?

I am currently conducting an experiment which involves FACS-sorting a specific population of cells using an antibody of interest.

In order to validate the type of cells I have collected using this marker, I want to perform qPCR on their extracted RNA, using certain known cell-type marker primers.

My problem is that in each sorting session, I am only able to collect anywhere from 5,000 - 10,000 cells. Do you know if it is possible to perform RT-qPCR on such a small sample size?

I have collected the cells in 1mL of Trizol.

Thanks for your help.