Nội dung được dịch bởi AI, chỉ mang tính chất tham khảo
Một chiến lược thêm vào hỗ trợ việc chọn lựa gen housekeeping cho phân tích RT-qPCR dưới các stress sinh học khác nhau ở quả cà tím
Tóm tắt
Các gen housekeeping nội sinh truyền thống được sử dụng để chuẩn hóa sự biểu hiện của các gen mục tiêu trong các nghiên cứu RT-qPCR. Giả định rằng không tồn tại một gen housekeeping nào hoàn hảo cho mọi điều kiện, sự ổn định biểu hiện của gen tham chiếu được chọn cần được đánh giá trong từng thí nghiệm mới. Quy trình chọn lựa gen housekeeping trở nên phức tạp và tốn thời gian hơn khi phải so sánh các điều kiện khác nhau trong cùng một tập dữ liệu thí nghiệm. Như một chiến lược thay thế, chúng tôi đã thêm vào một transcrit tham chiếu ngoại lai (ERT) vào tất cả các mẫu RNA trong tập dữ liệu của chúng tôi (rễ cà tím chịu các stress sinh học khác nhau), và sử dụng nó để chuẩn hóa mức độ biểu hiện của các gen housekeeping ứng cử viên nội sinh. Sự biểu hiện của ERT cho thấy sự ổn định cao trên tất cả các mẫu và cho phép chỉ ra glyceraldehyde 3-phosphate dehydrogenase (GAPDH) là gen housekeeping nội sinh ổn định nhất. Kết quả này đã được xác nhận thông qua việc sử dụng các thuật toán GeNorm, Normfinder và BestKeeper. Phương pháp này có thể được áp dụng một cách chung để rút ngắn quy trình chọn lựa gen tham chiếu tốt nhất.
Từ khóa
#gen housekeeping #RT-qPCR #cà tím #stress sinh học #transcrit tham chiếu ngoại lai #GAPDH #ổn định biểu hiện.Tài liệu tham khảo
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