CDKN1A exon 2, CDKN2B exon 2, and CDKN2A exons 1 2 were amplified under the same PCR conditions: in a touchdown PCR procedure the temperature of the reaction was lowered by 1°C every second cycle from 68°C to 60°C, at which temperature 30 cycles were carried out.
Standards were prepared with a 10-fold serial dilution (10-4 to 10 pg) of the PCR products and were run under the same PCR conditions used for the samples.
The 715 primer pairs were tested on wheat, rice, maize, cotton, and soybean under the same PCR conditions, and the effective primer pairs in the five crops were 500 (69.93%), 383 (53.57%), 452 (63.22%), 357 (49.93%), and 388 (56.27%), respectively.
The specificity of all these primers and two known primer sets for S. Typhimurium and S. Enteritidis was then confirmed under the same PCR conditions using 57 target strains and 112 nontarget Salmonella strains as well as 103 non-Salmonella strains.
One specific fragment (5.2 kb in size, Figure S1C, lane 3) was amplified using IS6100-R1, while no specific fragment was amplified under the same PCR conditions using IS6100-F1 (Figure S1C, lane 2).
Between eight and 30 recombinant clones per individual were amplified with primers OMNI (5'-ACA GGA AAC AGC TAT GAC CAT Gand3') and UNI (5'-CGA CGT TGT AAA ACG AGG CCA GT-3') under the same PCR conditions described above, and sized on 1.5% agarose gel.
miRNAs were subsequently amplified using 1 μl of the reverse transcribed sample, miRNA-specific forward and poly(T) adapter-specific reverse primers (additional file 6) under the same PCR-cycler conditions used in sqRT-PCR described above.
All PCR amplifications were carried out under identical conditions, on the same PCR plate and during the same reaction (β-actin was used as an internal control).
For all PCRs the same PCR conditions were used.
The same PCR conditions were used.
This potential weakness is amended in PLSR, as the extraction of components in PLSR aims to maximize the covariance with the outcome y: (A-4) under the same constraints for PCR that | w i | = 1 and w i ⊥ w j (i ≠ j) [ 44- 46].
