Journal: Molecular Reproduction and Development
Article Title: Identification and Migration of Primordial Germ Cells in Atlantic Salmon, Salmo salar: Characterization of Vasa, Dead End, and Lymphocyte Antigen 75 Genes
Figure Lengend Snippet: Distributionof vasa , dnd , and ly75 transcripts in Atlantic salmon. A : cDNA from various tissues of adult fish (blood, brain, gill, skeletal muscle, heart, liver, spleen, gall bladder, stomach, pyloric caeca, mid gut, head kidney, kidney, skin, testis, and ovary) were used for semi-quantitative RT-PCR. Actb was used as endogenous reference. Amplicon sizes, in base pairs, are indicated on the right. Expression pattern was determined using two biological replicates. B : Total RNA (400–900 ng) from early embryonic stages (two-cell, eight-cell, early-blastula, late-blastula, mid-gastrula, and 10-somite) was electrophoresed. Both 28S and 18S rRNA, stained with SYBR Safe DNA gel stain, are shown in all stages. C : The changes of both total RNA (white squares) and mRNA (black bars) amount per egg for each developmental stage. The concentration was quantified using three replicates. D : cDNA synthesized from above-mentioned developmental stages were used for semi-quantitative RT-PCR. In order to eliminate a possibility of genomic DNA contamination, −RT (without reverse transcriptase) samples of each counterpart were examined and electrophoresed. Amplicon sizes, in base pairs are indicated on the right.
Article Snippet: Total RNA and purified mRNA were electrophoresed on a 1% (w/v) agarose gel to assessed RNA integrity, and were further quantified with a NanoDrop ND-1000 (Thermo Scientific, Saven & Werner AS, Kristiansand, Norway).
Techniques: Fluorescence In Situ Hybridization, Quantitative RT-PCR, Amplification, Expressing, Staining, Concentration Assay, Synthesized