We generated 333-nucleotide (nt) sequences from the 70 samples for which qualitatively satisfactory sequences were obtained; all were identical.
In our experiments, ITS2, psbA-trnH, rpoC1, and rbcL provided not bad PCR efficiency (80% 96%) and not satisfactory sequencing efficiency (63%–73%), because AT-rich or homologous sequences existed, or concentration of PCR products was not high enough.
This ability would give the user the option of using highly stringent similarity requirements, the kind of requirements that would identify very few satisfactory sequences.
For weak PCR products resulting in non-satisfactory sequence quality, the reaction products were cloned into the pCR 2.1 TOPO vector (Invitrogen) for individual analysis.
Fluorescent sequencing was initially performed on the 104-bp ER-α exon 4 fragment that was used in SSCP and P-cycle sequencing, but this small amplicon did not yield satisfactory fluorescent sequencing data because of its high G/C content and the close proximity of primers to the codon of interest.
In our simulation, the sequencing depths (SD) of the four samples varied from 10 to 10. From Figure 7, we can see that the performances of FHB, FHC, and exomePeak are all satisfactory when sequencing depth is high enough (SD = 10); their performance all decreases together with the sequencing depth.
Using satisfactory spraying sequences (detailed herein), a reduction in the steel reinforcement of the RC layer can be obtained in every structural configuration and construction sequence, reaching a maximum percentage reduction of 7.0% of the total bending reinforcement.
Only samples that rendered both satisfactory DNA sequences and AFLP fingerprints were used for phylogenetic analysis.
Samples that did not produce a satisfactory whole-length sequence with these primers were amplified for the same region with four primer pairs producing shorter, overlapping, ~140-180 bproductsts (Additional file 11: Table S4).
After a detailed study, results indicate that our artificial swarm intelligence approach obtains satisfactory reliable DNA sequences.
Although there are several satisfactory alternative subject sequences, students who by the end of their second year have taken 14.01 Principles of Microeconomics and 14.02 Principles of Macroeconomics can follow a program that permits considerable depth in electives in their third and fourth years.
