For each of the nine patients, two sets of nested or semi-nested primers encompassing the fusion region were generated and used in a nested PCR reaction to amplify the fusion region directly from the patients' Guthrie cards (Andersen et al, 2001).
For sake of sensitivity and specificity, measurements were done by two-step quantitative RT-PCR, using two sets of nested primers annealing within the first Emx2 intron (Fig. 1A).
We used two sets of nested regression models to adjust for the confounding risk factors.
The cytochrome b gene (Cyt b) was amplified using two sets of nested primers.
To analyze genetic variation in HBoV2, two sets of nested primers were designed and used to amplify part of the NP1 and VP1 genes in positive samples (Table 1), using the NP1-F1 and NP1-R1, and VP1-F1 and VP1-R1 priners in the first round of PCR and the NP1-F2 and NP1-R2, and VP1-F2 and VP1-R2 in the second.
As our results from LRTs of all three sets of nested models on the DQA were highly significant, including M7 versus M8, we conclude that our conclusions hold up even under the low likelihood of undetected recombination.
Three sets of nested models were tested: 1) covariates (PCs, sex, age, ancestry by sex interaction), 2) covariates, SNP-GRSS and interaction with sex and 3) covariates, SNP-GRSS and three obesity-associated CNVs (the 21 kb deletion on 16p.12.3, the 66 kb duplication on 1p36.1, and the 1440 kb deletion on 5q13.2).
PCR amplification was carried out for HIV gag gene using two sets of primers in a nested strategy.
This is achieved through a difference in melting temperature between the two sets of primers used for the nested PCR such that the participation of each primer set can be specifically controlled by annealing temperature.
