Method for studying lin28 in regulating Blimp1 expression in process of differentiation from ESCs to PGCs of chickens

[0013] 1. Lin28A / B targeted gga-let7s screening

[0014] Using the online software http: / / mirdb.org / cgi-bin / search.cgi to predict chicken micRNA Let7s, a total of 17 gga-let7s were screened.

[0015] DF1 cells, ESCs and PGCs in good growth state were respectively selected and divided into 3 groups, one group was transfected with siLin28A / B, one group was transfected with oeLin28A / B, and the other group was not treated. DF1 cells were transfected in complete medium, and ESCs and PGCs were transfected in factor medium. At 37°C, 5% CO 2 After culturing for 48 h under saturated humidity, samples were taken to extract total RNA.

[0016] Total RNA was extracted according to the operating procedures of the miRNA extraction and isolation kit, cDNA was synthesized by reverse transcription according to the operating procedures of the reverse transcription kit, quantitative primers for micRNA-let7s were designed, and U6 was used as an internal reference to detect the relati