Your friend wants to develop a new method to map the transcriptionstart and end sites of a target gene because S1 mapping does notwork well. His professor suggests starting with the cDNA synthesismethod shown below. The reverse transcriptase has terminaltransferase activity and adds several cytosines to the end of firststrand cDNA. A template switch oligo with guanidines binds to thecytosines of cDNA and serve as new template. Your friend does notunderstand how a cDNA synthesis method can be used for transcriptstart and end sites mapping and asks you for help.LNA-containing TSOGrG+GCCCPoly(A)* RNAAAAAAAAAAAOligo(dT) primerReverse transcriptionand terminal transferaseAAAAAAAAAAQ4: As a hint for transcription start site mapping, his professorfurther suggests using a gene specific primer for reversetranscription instead of oligo(dT) primer. Please finish the logicaldesign of the method and briefly explain how the method works.Q5: To use the template switch mechanism for transcription end sitemapping, how will you achieve target gene specificity? Please finishthe logical design of the method and briefly explain how the methodworks.

Name: Your friend wants to develop a new method to map the transcriptionsta
Uploaded: 2024-03-20T06:42:41.000Z
Channel: Bartleby
Description: Your friend wants to develop a new method to map the transcriptionstart and end sites of a target gene because S1 mapping does notwork well. His professor suggests starting with the cDNA synthesismethod shown below. The reverse transcriptase has ter