Quantifiable RNA from invitro transcriptions?

Basavareju Shankarappa bsh at MED.PITT.EDU
Fri Jun 19 10:53:10 EST 1992


	Hi Netmeisters and meistresses:

	Can anyone tell me whether 
1.  We can obtain micro gram quantities of RNA using T3 or T7 transcription
reactions on PCR amplified or plasmid templates.  Or can we atleast obtain
RNA in quantities that can be quantitated with reasonable accuracy.

2.  What would be the quality of these RNAs?  If the template is about 250 base
long, will we have more than 90% (?) of full length transcripts.

	We need this information to obtain RNA for standardizing the 
sensitivity limit of the PCR reactions.  i.e., we need to know what is the
minimum copy number that we can amplify with our primers.  
	We would greatly appreciate any information you would have on this.
If there are any publications that you may want to guide us, it would also be
appreciated.

	Thanking you all in advance.  

Raj Shankarappa
Univ of Pittsburgh.



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