Microsatellite instability (MSI) analysis, carried out using a pentaplex panel of monomorphic mononucleotide repeats (BAT25, BAT26, NR-21, NR-22 and NR-24) as previously reported [ 12], demonstrated the absence of MSI in all tumor samples.
Microsatellite instability was determined in tumor DNA without the need for matching normal tissue DNA [ 23] using the highly specific panel of five quasimonomorphic mononucleotide repeats NR-21, NR-22, NR-24, BAT-25, and BAT-26 [ 23, 24] in a pentaplex polymerase chain reaction as previously described [ 25].
Using a highly sensitive pentaplex polymerase chain reaction to establish MSI status, cases were divided into microsatellite stable (MSS), MSI-low (MSI-L, 1 marker positive) and MSI-high (MSI-H, 2 5 markers positive).
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).
Kary Mullis came up with the idea in 1983, first using a polymerase from ordinary E. coli bacteria, but a polymerase was needed that could survive near-boiling temperatures.
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.
DNA of some of the female pup's genes had been replicated using a process called polymerase chain reaction.
APOE genotyping was conducted using a polymerase chain reaction-based assay.
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).
