isothermal amplification buffer ii pack  (New England Biolabs)

Journal: Scientific Reports

Article Title: Delay in primordial germ cell migration in adamts9 knockout zebrafish

doi: 10.1038/s41598-021-88024-x

Figure Lengend Snippet: Strong adamts9 expression in preovulatory follicular cells and developing embryos. ( A ) Expression of adamts9 determined by RT-PCR. Top panel is adamts9 transcript analyzed by RT-PCR (35 cycles), bottom panel is expression of a house keep gene, eukaryotic translation elongation factor 1 alpha 1a ( eef1a1a ). M. NEB 1 kb plus DNA ladder; 1. Ovary (fully grown immature follicles); 2. Preovulatory stage IVb follicles; 3. Ovulated stage V oocytes; 4. One cell stage embryos; 5. Four cell stage embryos; 6. Oblong stage embryos (~ 3.5 hpf, hours post fertilization); 7. Germ-ring stage embryos (~ 5.5 hpf); 8. Eight somite stage embryos (~ 11.5 hpf); 9. 24 hpf (hours post fertilization) embryos; 10. Two dpf (days post fertilization ) embryos; 11. Three dpf embryos; 12. Six dpf embryos; 13. Six wpf (weeks post fertilization) gonad. Please see Supplemental Figs. 3 and 4 for full agarose gel images. ( B ) Strong adamts9 expression in preovulatory follicles, low or no expression in immature follicles or ovulated oocytes determined by real-time quantitative PCR (qPCR). Stage I and III immature follicles, IVb preovulatory follicular cell enclosed oocytes and stage V ovulated oocytes were collected from mature AB wildtype females between 7–8:30 am (lights on 8:30am-10:30 pm). Results were presented as mean ± SEM (n = 5). Different letters above the error bars indicate that those groups are significantly different from each other at p

Article Snippet: A set of PCR primers targeting 242 bp of coding region of eef1a1a (forward: 5′-AGTGTTGCCTTCGTCCCAAT-3′; Reverse:5′-CACACGACCCACAGGTACAG-3′) was used for PCR amplification.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction