TLS Online TPP Program

#Id: 2632

#Unit 13. Methods in Biology

The principle behind the S1 mapping method is to label a single stranded DNA probe that can hybridize only to the transcript of interest. 

TLS Online TPP Program

#Id: 2633

#Unit 13. Methods in Biology

The probe must span the sequence where the transcript starts or ends. After hybridizing the probe to the transcript, one applies S1 nuclease, which degrades only single-stranded DNA and RNA. 

TLS Online TPP Program

#Id: 2634

#Unit 13. Methods in Biology

The probe must span the sequence where the transcript starts or ends. After hybridizing the probe to the transcript, one applies S1 nuclease, which degrades only single-stranded DNA and RNA.

TLS Online TPP Program

#Id: 2635

#Unit 13. Methods in Biology

Double-stranded DNAs, RNAs, and hybrids are protected from S1 nuclease degradation. 

TLS Online TPP Program

#Id: 2636

#Unit 13. Methods in Biology

S1 mapping the 5’-end of a transcript

TLS Online TPP Program

#Id: 2637

#Unit 13. Methods in Biology

Primer extension Using primer extension one can locate the 59-end of a transcript by hybridizing an oligonucleotide primer to the RNA of interest, extending the primer with reverse transcriptase to the 5’-end of the transcript, and electrophoresing the reverse transcript to determine its size