3 μl of template were used in the first round of PCR, using primer C5 coupled with either primer 138 or NS8 (Table 1). 1 μl of product from the first round was used as the template in the second round, initially using primers 2082F and 2514R (Table 1) for the identification of genetic types.
The first round of PCR uses gene specific primers while the second round is used to incorporate the dU containing primers used to create the complementary ss-tails.
cDNA was used as a template in a first round of PCR using specific primers corresponding to exon 1 of STAG2 (5′-GAGGGAACAACATTCATGTG-3′) and the carp β-actin splice acceptor sequence (5′-CATACCGGCTACGTTGCTAA-3′).
For embryos derived from Pdcd2 +/− and Pdcd2 +/lacZ intercrosses, the first round of PCR used a mixture of LAR3, 3′arm and 5′arm primers and the second round of PCR used a mixture of Nested LAR3, Nested 3′arm and Nested 5′arm primers (supplementary material Table S7).
The first round of PCR used the linker specific primer and a downstream gene specific primer, while the second round used a portion of the first round product and a gene specific primer 3' to that of the first round gene specific primer to increase specificity of the reaction.
The first round of coding used the pre-identified themes as written into the interview schedule.
The first round of reactions used two of these sets to detect Salmonella O 4, O 9, O 7, O 8, and O 3,10 serogroups.
A first round of PCR used primers generated by Primer3 [29] to amplify from genomic DNA samples.
Second, we employed a straightforward optimization protocol for the sequence length cut-off, starting with a first round of calculations using the lengths L = 10, 20, 30, 40 and 50 only.
The first round of PCR used P1 and P2 primers.
