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TLS Online TPP Program

#Id: 6105

In contrast to most alkylating agents, the hydroxylating agent hydroxylamine (NH2OH) has a specific mutagenic effect. It induces only G:C → A:T transitions.


#Unit 3. Fundamental Processes #Mutation and Repair #Part B Pointers
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TLS Online TPP Program

#Id: 2640

#Unit 13. Methods in Biology

Run-off transcription is a means of checking the efficiency and accuracy of in vitro transcription.
TLS Online TPP Program

#Id: 2641

#Unit 13. Methods in Biology

The size of this run-off transcript locates the transcription start site, and the amount of this transcript reflects the efficiency of transcription. 
TLS Online TPP Program

#Id: 2642

#Unit 13. Methods in Biology

Notice that S1 mapping and primer extension are well suited to mapping transcripts made in vivo; by contrast, run-off transcription relies on transcription in vitro.
TLS Online TPP Program

#Id: 2643

#Unit 13. Methods in Biology

After you have identified the physiological transcription start site by S1 mapping or primer extension you can use run-off transcription in vitro.
TLS Online TPP Program

#Id: 2644

#Unit 13. Methods in Biology

A variation on the run-off theme of quantifying accurate transcription in vitro is the G-less cassette assay.
TLS Online TPP Program

#Id: 2645

#Unit 13. Methods in Biology

In G-less cassette transcription, a promoter is fused to a double stranded DNA cassette lacking G’s in the nontemplate strand, then the construct is transcribed in vitro in the absence of GTP.