【貨號】 BC-CL01【規(guī)格】 1ml/1.5ml
【保存】 2-8℃，12個月
【類型】 陽離子聚合物
【產(chǎn)品介紹】
BC Trans-DNA plus是一款專為難以轉(zhuǎn)染的細胞設(shè)計的DNA轉(zhuǎn)染試劑，它能夠高效地傳送質(zhì)粒DNA。相較于其他轉(zhuǎn)染試劑，BC Trans-DNA plus擁有卓越的DNA轉(zhuǎn)染能力，其優(yōu)勢在于高轉(zhuǎn)染效率、血清干擾小、低毒性、良好穩(wěn)定性、操作簡便以及出色的可重復性。
【應用范圍】
BC Trans-DNA plus轉(zhuǎn)染試劑表現(xiàn)出廣泛的適用性，能夠高效應對多種難轉(zhuǎn)染細胞株的DNA轉(zhuǎn)染需求，無論是瞬時轉(zhuǎn)染還是穩(wěn)定轉(zhuǎn)染。特別值得一提的是，該試劑在多種貼壁細胞系中表現(xiàn)出色，尤其對于BMDC、4T1、CT26、DC2.4等難以轉(zhuǎn)染的細胞類型，依然能夠?qū)崿F(xiàn)高轉(zhuǎn)染效率，并且具備良好的重復性。
【質(zhì)粒DNA轉(zhuǎn)染步驟】
請確保所有操作都在無菌條件下進行，并遵循實驗室的安全規(guī)范。
以24孔板為例，請參考表1的轉(zhuǎn)染規(guī)模調(diào)整，步驟如下:
1. 細胞接種：在細胞培養(yǎng)板中，每孔接種0.5~1.0×105個細胞。將細胞在適宜條件下培養(yǎng)12~24小時，以確保在轉(zhuǎn)染時細胞密度達到約60~70%的融合度。
2. DNA稀釋：將所需的質(zhì)粒DNA用Opti-MEM培養(yǎng)基稀釋，使其終濃度達到0.1mg/mL。
3. 復合物制備：
（1）取1µL的BC Trans-DNA plus試劑原液，加入9µL的Opti-MEM培養(yǎng)基中。
（2）同時，取4µL濃度為0.1mg/mL的質(zhì)粒DNA，加入6µL的Opti-MEM培養(yǎng)基中。
（3）將上述兩者等體積混合，以形成轉(zhuǎn)染復合物。
（4）室溫靜置：讓轉(zhuǎn)染復合物在室溫下靜置10分鐘，以確保其充分結(jié)合。
4. 轉(zhuǎn)染細胞：將20µL的轉(zhuǎn)染復合物均勻加入到細胞培養(yǎng)板的每個孔中，并輕輕混勻。
5. 培養(yǎng)與檢測：將細胞在37℃下繼續(xù)培養(yǎng)18~48小時。在此期間，無需更換培養(yǎng)基。
【質(zhì)粒DNA的轉(zhuǎn)染優(yōu)化】
可通過改變細胞密度、 DNA濃度以及BC Trans-DNA plus濃度對轉(zhuǎn)染進行優(yōu)化。保證細胞融合度在60%以上，BC Trans-DNA plus (µL):DNA (µg)可以在2:1和5:1之間調(diào)整。
表1. 不同培養(yǎng)板所需轉(zhuǎn)染試劑和DNA的用量
| 培養(yǎng)板 | | 接種 培養(yǎng)基 | Opti-MEM稀 釋后終體積 | DNA轉(zhuǎn)染 |
| 試劑用量 | DNA |
| 96孔板 | 0.3cm2 | 200µL | 10µL | 0.5µL | 0.2µg |
| 24孔板 | 2.0cm2 | 500µL | 20µL | 1µL | 0.4µg |
| 12孔板 | 4.0cm2 | 1mL | 40µL | 2.5µL | 1µg |
| 6孔板 | 10.0cm2 | 2mL | 100µL | 5µL | 2µg |
| 60mm | 20.0cm2 | 5mL | 0.2mL | 10µL | 4.0µg |
| 10cm | 60.0cm2 | 15mL | 0.6mL | 30µL | 12µg |
標簽：細胞轉(zhuǎn)染
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