Primary granulosa cells treated with 5×10 10 PFU/ml adv-control or adv-lnc-GULP1-2:1 (Hanbio Biotechnology, Shanghai, China) for 48h were seeded (70% con uent) in 12-well plates for ow cytometry analysis. Cells were digested with EDTA-free trypsin, washed twice with cold PBS (1000 rpm × 5 min), and then xed with 70% ethanol at 4 ℃ overnight. The xed cells were centrifuged (1000 rpm × 5 min) again on the next day, resuspended in 500μl of PI/RNase solution KeyGen Biotech Nanjing China), and then incubated in the dark for 30 minutes at 37 ℃. Flow cytometry studies were performed by using BD Accuri C6 Plus ow cytometer (BD Biosciences, San Jose, CA). The percentage of cells in different phases of cell cycle were analyzed by using FlowJo 10.4 (BD Biosciences).
Pi rnase solution
Manufactured by Keygen Biotech
Sourced in China
The PI/RNase solution is a laboratory reagent designed for use in various molecular biology applications. It contains a combination of proteinase inhibitors (PI) and RNase enzymes, which serve to protect nucleic acids from degradation during sample processing and analysis.
Automatically generated - may contain errors
3 protocols using pi rnase solution
1
Adv-lnc-GULP1-2 Regulates Granulosa Cell Cycle
2
Cell Cycle Analysis of Primary Granulosa Cells
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Primary granulosa cells treated with 5 × 1010 PFU/ml adv-control or adv-lnc-GULP1–2:1 (Hanbio Biotechnology, Shanghai, China) for 48 h were seeded (70% confluent) in 12-well plates for flow cytometry analysis. Cells were digested with EDTA-free trypsin, washed twice with cold PBS (1000 rpm × 5 min), and then fixed with 70% ethanol at 4 °C overnight. The fixed cells were centrifuged (1000 rpm × 5 min) again on the next day, resuspended in 500 μl of PI/RNase solution (KeyGen Biotech, Nanjing, China), and then incubated in the dark for 30 min at 37 °C. Flow cytometry studies were performed by using BD Accuri C6 Plus flow cytometer (BD Biosciences, San Jose, CA). The percentage of cells in different phases of cell cycle were analyzed by using FlowJo 10.4 (BD Biosciences).
3
Adenoviral Regulation of Granulosa Cell Cycle
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Primary granulosa cells treated with 5×10 10 PFU/ml adv-control or adv-lnc-GULP1-2:1 (Hanbio Biotechnology, Shanghai, China) for 48h were seeded (70% con uent) in 12-well plates for ow cytometry analysis. Cells were digested with EDTA-free trypsin, washed twice with cold PBS (1000 rpm × 5 min) and then xed with 70% ethanol at 4 ℃ overnight. The xed cells were centrifuged (1000 rpm × 5 min) again on the next day, resuspended in 500μl of PI/RNase solution KeyGen Biotech Nanjing China), and then incubated in the dark for 30 minutes at 37 ℃. Flow cytometry studies were performed by using BD Accuri C6 Plus ow cytometer (BD Biosciences, San Jose, CA). The percentage of cells in different phases of cell cycle were analyzed by using FlowJo 10.4 (BD Biosciences).
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