Practice question

Radioactive labeling of primers for detection in Sanger sequencing, footprinting, or probe tracking is best achieved by transferring γ-phosphate of [γ-32P]ATP to 5' hydroxyl terminus of oligonucleotide using T4 polynucleotide kinase, yielding 5' 32P-labeled primer. This end-label does not interfere with polymerase extension from 3' end. Alpha-32P labeled deoxynucleotides incorporate internally via polymerase synthesis but not suitable for pre-synthesized primer labeling. Beta and generic notations are not used. Five-prime labeling provides high specific activity enabling autoradiographic detection with superior sensitivity.