【貨號(hào)】 BC-CL03【規(guī)格】 1ml/1.5ml
【保存】 2-8℃，12個(gè)月
【類型】 陽離子聚合物
【產(chǎn)品介紹】
BC Trans-mRNA是一款高性能的專用轉(zhuǎn)染試劑，專為mRNA設(shè)計(jì)，適用于在多種貼壁細(xì)胞中進(jìn)行mRNA轉(zhuǎn)染。這款試劑的獨(dú)特之處在于，它能夠直接將mRNA傳遞到細(xì)胞質(zhì)中進(jìn)行表達(dá)，從而巧妙地繞過了轉(zhuǎn)錄調(diào)控作用以及進(jìn)入細(xì)胞核的限制。因此，BC Trans-mRNA成為短期蛋白表達(dá)相關(guān)研究工作的理想選擇，為科研人員提供了便捷、高效的工具。
【應(yīng)用范圍】
BC Trans-mRNA轉(zhuǎn)染試劑展現(xiàn)出廣泛的適用性，能夠高效應(yīng)對(duì)眾多貼壁或懸浮細(xì)胞株的mRNA轉(zhuǎn)染需求。無論是在常規(guī)細(xì)胞、難以轉(zhuǎn)染的細(xì)胞、原代細(xì)胞還是干細(xì)胞中，該試劑都能實(shí)現(xiàn)較高的轉(zhuǎn)染效率，并且性能穩(wěn)定可靠。與其他轉(zhuǎn)染試劑相比，BC Trans-mRNA對(duì)血清的干擾極小、毒性低、穩(wěn)定性好、操作簡(jiǎn)便易行以及良好的重復(fù)性。
【mRNA的轉(zhuǎn)染步驟】
以24孔板為例，請(qǐng)參考表1的轉(zhuǎn)染規(guī)模調(diào)整，步驟如下:
1. 細(xì)胞接種: 每孔接種0.5~1.0×105個(gè)細(xì)胞，細(xì)胞培養(yǎng)12~24小時(shí)，使轉(zhuǎn)染時(shí)細(xì)胞密度達(dá)到60~70%融合度。
2. mRNA稀釋: 將mRNA稀釋于Opti-MEM培養(yǎng)基中，終濃度0.1mg / mL。
3. 復(fù)合物制備:
（1）取1µL的BC Trans-mRNA試劑原液加入到9µL Opti-MEM培養(yǎng)基中
（2）取4µL濃度為0.1mg / mL的mRNA加入到6µL Opti-MEM培養(yǎng)基中
（3）兩者等體積混合
（4）室溫靜置10分鐘
4. 轉(zhuǎn)染細(xì)胞：將20µL的轉(zhuǎn)染復(fù)合物均勻加入到細(xì)胞培養(yǎng)板的每個(gè)孔中，并輕輕混勻。
5. 培養(yǎng)與檢測(cè)：將細(xì)胞在37℃下繼續(xù)培養(yǎng)18~48小時(shí)。在此期間，無需更換培養(yǎng)基。
【mRNA的轉(zhuǎn)染優(yōu)化】
可通過改變細(xì)胞密度、 mRNA濃度以及BC Trans-mRNA濃度對(duì)轉(zhuǎn)染進(jìn)行優(yōu)化。保證細(xì)胞融合度在60%以上，BC Trans-mRNA (µL): mRNA (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mRNA的用量
| 培養(yǎng)板 | | 接種 培養(yǎng)基 | Opti-MEM培養(yǎng) 基釋后終體積 | mRNA轉(zhuǎn)染 |
| 試劑用量 | mRNA |
| 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 | 200µL | 10µL | 4.0µg |
| 10cm | 60.0cm2 | 15mL | 600µL | 30µL | 12µg |
標(biāo)簽：轉(zhuǎn)染
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20、Bo Qiu,Ming Gong,Qi-Ting He,etc.Controlled Release of Interleukin-1 Receptor Antagonist from Hyaluronic Acid-Chitosan Microspheres Attenuates Interleukin-1β-Induced Inflammation and Apoptosis in Chondrocytes[J].Biomed Res Int.2016:1-12.
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