주문정보
- - 재고수량은 변동될 수 있습니다.
- - 재고 확인 시 "갱신" 버튼을 누르시면 실시간 재고를 확인하실 수 있습니다.
- - 가격이 ‘별도문의‘ 시, 상단 ‘견적신청’ 버튼을 눌러 문의해주시면 빠른 답변을 받으실 수 있습니다.
| 선택 | Cat.No. | 제품명 | 가격(VAT별도) | 수량 |
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제품특징
□ 특징
● CRISPR/Cas9을 위한 sgRNA 및 mRNA, miRNA, siRNA등 여러 종류의 RNA Transfection에 최적
● 다양한 종류의 포유류 세포 (mammalian cell)에적용 가능
● 높은 효율의 유전자 도입과 매우 낮은 세포 독성
● Simple, serum-compatible protocol
● Transfection에 필요한 모든 구성품을 포함한 All-in-One system
□ 제품설명
Xfect™ RNA Transfection reagent는 다양한 종류의 mammalian cell (broad-range)에microRNA, siRNA, mRNA 또는 sgRNA와같은 여러 종류의 RNA를 효율적으로 도입 가능한Transfection 시약이다. 생분해성 나노입자 (biodegradablenanoparticles)로 구성된 Xfect™ Transfection reagnet는 일반적인 Transfection 시약에 비하여 낮은 세포독성및 높은 도입효율로 RNA Transfection이 가능하다. 특히 본 제품은 CRISPR/Cas9-mediated gene editing에 사용되는Cas9 coding mRNA 및 single guide RNA 도입에 효율적으로 사용이가능하며, Xfect™ Transfection reagent(Code 631317, 631318)과 함께 사용하면, DNA와 RNA co-transfection이 가능하다.
그림 1. Xfect™ RNA Transfection 시약을 이용한 sgRNA Transfection 후,CD81 gene knock-out 결과 Panel A. sgRNA targeting the 5’ end of the antisensestrand of CD81 was synthesized using the Guide-it sgRNA In Vitro Transcription Kit. Panel B. An HT1080 cell line (2.0 x105 cells) stably expressing Cas9 (HT1080-Cas9) was transfectedwith 50 pmol of sgRNA targeting CD81, either once or twice (lowergraph), using the Xfect RNA Transfection Reagent. Seven days later, cells wereimmunostained with a CD81 antibody (Ab) conjugated to an FITC fluorophore andanalyzed by flow cytometry. The percentage of cells that did not bind CD81 wascalculated. A control sample, comprised of HT1080-Cas9 cells, was analyzed byflow cytometry, either without (top, left graph) or with (top, right graph) theCD81 antibody. Both single and double transfection with sgRNA resulted in asubstantial increase in cells that did not bind CD81, indicating successfulCRISPR/Cas9-mediated knockout of CD81.
그림 2. Primarycell 및 다양한 종류의 cell line에서의mRNA transfection 효과 HeLa cells (2.0 x105), HEK 293 cells (1.5 x 105), Human Dermal Fibroblasts(NHDF) cells (6.0 x 104), Mesenchymal Stem Cells (MSCs) (5.0 x 104),Jurkat Cells (3.0 x 105), and KBM7 cells (3.0 x 105) wereplated in 12-well plates and transfected with 1 μg of mRNA encoding GFP with 5μl of Xfect RNA Transfection Reagent. 20 hours later, cells were analyzed byflow cytometry and the % GFP-positive cells and the Mean Fluorescence Intensity(MFI) were determined.
그림 3.Primary cell 및 cell line에서 siRNATransfection에 의한 Knock-down 효율HeLa cells (2.0 x 105),Human Dermal Fibroblasts (NHDF) cells (6.0 x 104), and MesenchymalStem Cells (MSCs) (5.0 x 104)were plated in 12-well plates andtransfected with 50 pmol of siRNA against luciferase using Xfect RNATransfection Reagent.All three cell types were also transfected with 1 μg ofpCMV-Luc using the Xfect Transfection Reagent.Luciferase assays were performed 48 hourspost-transfection. For control samples, cells were transfected with pCMV-Lucbut without the siRNA.We observed a dramatic (>95%) decrease in luciferaseactivity in all the cells treated with siRNA.
□ Applications
● CRISPR/Cas9
● RNAtransfection
● Transienttransfection without genomic integration
□ 구성품
Xfect RNATransfection Polymer | 600ul× 2 |
Xfect ReactionBuffer | 12ml× 2 |
□ 보존 - 20 ℃
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1、根据所需求的酸碱性选择合适的缓冲对，若是配制酸性缓冲液就选择弱酸与弱酸盐缓冲对；若是配制碱性缓冲液就选择弱碱与弱碱盐缓冲对。
2、根据所需要控制的PH范围以及弱酸和弱碱的解离常数（pKa/pKb）选择具体的共轭酸碱对，公式为PH＝pKa±1＝（14-pKb）±1。
3、根据公式计算缓冲溶液的组分比，酸性缓冲液PH＝pKa－lgc酸/c盐，碱性缓冲液PH＝14-pKb+lgc碱/c盐。
4、根据共轭酸碱对以及其组分比配制缓冲液，方法同普通溶液。
举例：试配制一种缓冲液，体积为1L，PH能维持在10.25左右。
a、依题意选择弱碱与弱碱盐的共轭对；
b、由PH＝（14-pKb）±1，算出pKb在3.75与4.75之间，查弱碱的解离常数表可知氨水（pKb=4.75）符合要求，故可选择NH3-NH4Cl体系；
c、由PH＝14-pKb+lgc碱/c盐，算出c(氨水)/c(氯化铵)＝10；
d、设氨水的浓度为10mol/L，则氯化铵的浓度为1mol/L，所以在浓度为10mol/L体积为1L的氨水中加入1mol的氯化铵即可。
酶提取技术，属地球化学勘查学科。其是由克拉克（J.R.Clark）等人于20世纪80年代末和90年代初研制出的一种利用葡萄糖氧化酶提取矿物颗粒表面的非晶质锰的氧化膜寻找隐伏矿的方法。1995年以后已广泛应用。
根据组成不同，可分为两种，弱酸及其对应的强碱弱酸盐，弱碱及其对应的强酸弱碱盐。
因为HF可以和NaOH反应生成NaF和水，当NaOH反应完之后，NaF就可以与HF组成缓冲溶液，所以说可以直接使用NaOH和HF来配制缓冲溶液。
这也是一般配制缓冲溶液的方法，也就是用强碱和弱酸（或者强酸和弱碱）来配制缓冲溶液。
