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The primer sequences used were: Cecropin G (AAEL015515-RA): F: 5'-TCACAAAGTTATTTCTCCTGATCG-3' R: 5'-GCTTTAGCCCCAGCTACAAC-3' Defensin C (AAEL003832-RA): F: 5'-TTGTTTGCTTCGTTGCTCTTT-3' R: 5'-ATCTCCTACACCGAACCCACT-3' S7 (AAEL009496-RA): F: 5'-GGGACAAATCGGCCAGGCTATC-3' R: 5'-TCGTGGACGCTTCTGCTTGTTG-3' Numeric data for the semi-quantitative PCR assays are presented in Table S3.
The mutational status of K-Ras (codon 12,13) was performed using 40 ng of genomic DNA amplified by PCR in 25 μl reaction mix containing 1.25 U Fast Start High Fidelity Taq (Roche), 0.2 m M dNTP, 1.5 m M MgCl2, and 0.2 μm of K-Ras F and K-Ras R I primers (Table 2).
GH, RA-F, GG, LG participated in drafting the manuscript.
RA-F, GG and GH coordinated the work and participated in analyzing results.
GG and RA-F participated in sequence and phylogenetic analyses, and Mapman analysis of microarray data.
There are at least six isoforms of the leptin receptor (OB-Ra-f), primarily as a consequence of alternate splicing.
Alternate splicing of the db gene generates multiple variants of Ob-R mRNA and at least six receptor isoforms (Ob-Ra-f) have been identified [ 8].
RA-F, ML, JA, MR, OVL, LG, GG, DP, BC, PC and FSp participated in the different experimental approaches used, such as: plant growth, plant transformation, determination of gene expression, plasmid construction, protein cellular localization, microarray hybridization and densitometry.
Western blot analyses were done on nitrocellulose membranes hybridized with various antibodies: from Santa Cruz p53 (DO-1, sc-126), p21 (F5, sc-6246), H-Ras (F-235, sc-29), p16 (JC-8, sc-56330), PCNA (FL-261, sc-7907), from Millipore (Upstate) Necdin (07 565) and from Abcam β-actin (AC-15, ab6276).
The velocity components are given by u = ( α m x ) Ra x f ′ and v = ( α m 2 x ) Ra x 1 2 [ y x Ra x 1 2 f ′ − f ]. (8).
Morey RA, Dolcos F, Petty CM, Cooper DA, Hayes JP, LaBar KS et al (2009).
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