cDNA amount - (Jun/13/2005 )
The protocol: "we used Sensiscript and Superscript II (GibcoBRL, Inchinnan, UK) and oligo-dT to prepare cDNA from 2,5 ng samples of total RNA isolated from a colorectal cancer cell line. The equivalent of 0,125 ng were used in qPCR assays. "
What does it mean with “The equivalent of 0,125 ng”? Is this purely cDNA or the reverse transcribed sample (cDNA+ initial RNA left)?
-indoubt-
QUOTE (indoubt @ Jun 13 2005, 09:58 AM)
The protocol: "we used Sensiscript and Superscript II (GibcoBRL, Inchinnan, UK) and oligo-dT to prepare cDNA from 2,5 ng samples of total RNA isolated from a colorectal cancer cell line. The equivalent of 0,125 ng were used in qPCR assays. "
What does it mean with “The equivalent of 0,125 ng”? Is this purely cDNA or the reverse transcribed sample (cDNA+ initial RNA left)?
What does it mean with “The equivalent of 0,125 ng”? Is this purely cDNA or the reverse transcribed sample (cDNA+ initial RNA left)?
If you do your RT step of 2.5ng RNA in a 20ul reaction. Then 1 ul of that reaction volume will be equivalent to .125ng of rna. In actuality you will be using 1ul of cdna that represents that .125ng of rna.
-dobbiewalton-