The 1.5 kbp products were subjected to Sanger dideoxy sequencing
using the forward primer and reverse primers at Macrogen DNA, Inc.
Both of these variants were flagged to have poor quality scores and would have been flagged by the laboratory for confirmation by Sanger dideoxy sequencing
Following preliminary population surveys via dideoxy sequencing
(Alter and Palumbi 2009), we designed two mtDNA markers for haplotyping.
Direct sequence analysis of PCR products was performed in both forward and reverse directions using automated fluorescence dideoxy sequencing
was performed to confirm the variant selected as potentially relevant to disease and test for genotype-phenotype cosegregation in the family.
However, requiring relatively low-throughput dideoxy sequencing
as a validation of high-throughput NGS interrogation severely limits the utility of NGS.
Sanger dideoxy sequencing
method was followed in which the primer: 518F-5'CCAGCAGCCGCGGTAAT ACG3' and 800R-5"TACCAGGGTATCTAATCC3' was used for amplification process.
Respiratory specimens from the 2 infected humans and the 5 dromedaries that were still positive at the second sampling were analyzed by dideoxy sequencing
as previously described (13); the nucleocapsid gene sequences of all dromedary samples were found to be identical.
reactions employed BigDye version 3.1 chemistry and a Prism 3730XL sequencer (Applied Biosystems, Foster City, CA, U.S.A.) using ~50-100 ng of PCR template.
Most traditional sequencing, or Sanger Dideoxy sequencing
, is performed on PCR products.
Sakaki, "Dideoxy sequencing
method using denatured plasmid templates," Analytical Biochemistry, vol.
Sanger Sequencing, also called Sanger Dideoxy Sequencing
, is the controlled interruption of enzymatic DNA replication through the use of dideoxynucleotides in primer-directed DNA extension to produce discrete DNA fragments (11,41) (Fig.