Method for obtaining FSHR full-length coding region sequence with multiple splice forms
A coding region and sequence technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of heavy workload, high test cost, high cost, etc., and achieve the effect of roughly clear length, highlighting technological progress, and low cost
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[0033] Step 1 extracts the total RNA (ribonucleic acid) in the sheep granulosa cells, and then reverse-transcribes the total RNA into cDNA;
[0034] 1. Collection of Tissue
[0035] Collect sheep ovaries from the slaughterhouse, keep them warm in sterilized normal saline at 30°C, and transport them to the laboratory within 2 hours. Extract granulosa cells and follicular fluid with a 10mL sterile syringe in the laboratory, and transfer the extracted follicular fluid and granulosa cells to a 1.5mL centrifuge. Centrifuge in the tube at 4°C, 4000rpm, 5min, discard the supernatant and save the precipitate, which is used for RNA extraction.
[0036] 2. RNA preparation and cDNA synthesis
[0037] Add 1.0mL Trizol solution to the 1.5mL EP tube precipitation, pump repeatedly 10-20 times with a 1mL sterilized syringe until the cell lysis solution is completely clear, and place at room temperature for 5min.
[0038] Extract sheep follicle fluid with a 10mL syringe needle, sort out oocy...
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