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Hoang TM, Kent T, Pomerantz RT
Modification of 3' Terminal Ends of DNA and RNA Using DNA Polymerase theta Terminal Transferase Activity
Bio Protoc (2017) In process.
DNA polymerase theta (Poltheta) is a promiscuous enzyme that is essential for the error-prone DNA double-strand break (DSB) repair pathway called alternative end-joining (alt-EJ). During this form of DSB repair, Poltheta performs terminal transferase activity at the 3' termini of resected DSBs via templated and non-templated nucleotide addition cycles. Since human Poltheta is able to modify the 3' terminal ends of both DNA and RNA with a wide array of large and diverse ribonucleotide and deoxyribonucleotide analogs, its terminal transferase activity is more useful for biotechnology applications than terminal deoxynucleotidyl transferase (TdT). Here, we present in detail simple methods by which purified human Poltheta is utilized to modify the 3' terminal ends of RNA and DNA for various applications in biotechnology and biomedical research.
Publication Date: 2017-06-20.
PMCID: PMC5560110
Last updated on Sunday, July 12, 2020