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In order to test our hypothesis and investigate the presence of intraisolate allelic variations within AMF mtDNA, we returned to the basics, using a reliable polymerase chain reaction (PCR -based aPCR -basedth approachidelity DNA polymerase, followithby cloning and Sanger sequencing of selected mtDNA regions.
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To obtain a reliable polymerase kinetic signal for 5mC, the DNA was treated with Tet1 oxidation before sequencing [ 28].
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The three selected BVMO-encoding genes; Af1 (XM_742067), Af2 (XM_741856) and Af3 (XM_750181) were amplified by PCR using a proofreading polymerase (Phusion DNA polymerase, NEB).
Dr. Constable then extracted the DNA from the stools and amplified them, using a technique called polymerase chain reaction, or P.C.R., the same method that Dr. Gagneux had applied to amplifying the hair DNA.
Genomic insertion was confirmed using a polymerase chain reaction-based method and Southern blots.
Gene expression was performed using a reverse transcriptase polymerase chain reaction (RT-PCR).
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