2×实时定量PCR扩增预混液

规格与询价

勾选需要的规格可一次提交；也可以逐个询价。货号会一并带入询价单。

货号规格价格状态

11184ES03 1 mL $383.00 现货询价

11184ES08 5× 1 mL $668.00促~~$1,653.00~~ 现货询价

11184ES50 50× 1 mL $12,533.00 现货询价

11184ES60 100× 1 mL $19,833.00 现货询价

提交询价价格为协议价，提交后由业务确认；大包装可申请试用装。

产品概述

技术参数

品牌翌圣生物 Yeasen

货号 / 型号11184ES

规格全仪器平台通用：预混特定类型参比染料，无需调整ROX浓度易于示踪：蓝色预混液，观察颜色可直接区分是否加样强稳定性：室温配制体系，配制后的体系可4℃放置24 h，检测结果无变化上机快速：兼容常规程序与快速程序，最快46 min完成定量实验扩增

产地翌圣生物科技(上海)股份有限公司

技术平台 实时荧光定量PCR 通用qPCR平台

应用方法 染料法荧光定量PCR基因表达定量分析

适用物种种属人

检测方法 SYBR Green染料法

生物特性参数 试剂类型荧光定量预混液（2×实时定量PCR预溶液） 应用染料法荧光定量PCR 灵敏度可检测至单拷贝 特异性高特异性 GC含量适用范围30-70% 线性检测范围7个数量级 分辨率可精准分辨2倍模板浓度差异 重复性90个复孔Ct值标准偏差<0.2

样本类型质粒 cDNA

技术参数 产品形式预混液 热启动方式抗体法热启动 ROX参比染料适用于绝大多数机型 扩增长度80-300bp 储存条件-20℃避光储存，有效期18个月，避免反复冻融 运输条件冰袋运输 仪器兼容性适用于所有qPCR仪器 反应时间最快46 min完成定量实验

实验类型 实时荧光定量PCR基因定量扩增

常见问题

Q：建议qPCR 实验用几步法？

A：常用 2 步法。需提高扩增特异性，可选用 2 步法或提高退火温度。在扩增效率低， ct 值过大的时候，可以改用 3 步法或延长延伸时间。

Q：qPCR 实验结果的有效性？为什么建议Ct 值要大于 15？

A：有效性要满足三个条件：（1）标准曲线：扩增效率范围:90-110%，对应斜率为 -3--3.5。 R2＞0.98。 (扩增效率=10-1/斜率-1)，当斜率=-3.32 时，扩增效率=100%。（2）扩增曲线：S 型曲线，且 Ct 值在 15－35 之间，阴性对照 Ct＞35 或无 Ct 值。（3）熔解曲线：为单一峰。

Ct 值大于 15 个循环是因为 3-15 个循环的荧光值标准差的 10 倍是荧光阈值，Ct 值太小了会影响曲线。

Q：11184 的灵敏度极限是?

A：单拷贝

Q：同一基因复孔间熔解曲线 Tm 值有差异？

A：同样的扩增产物也会出现Tm 值有微小差异，一般差异在 1 度以内都可以接受。

Q：为什么稀释了模板CT 值反而变小了？

A：一般 CT 值与模板起始浓度呈负相关，浓度越高，CT 值越小。但也有很多特殊情况，比如体系中存在抑制物或是模板不纯，这时候稀释模板反而能使 CT 值变低。

Q：内参CT 值小于 20，目的基因均大于 30，怎么办？

A:可能是目的基因为低丰度表达基因导致。建议：a）换用内参；b）换引物。

Q：为什么扩增曲线杂乱且不连续？

A：可能原因是 ROX 添加不当。确认参比染料ROX 是否添加正确。

Q：模板用量X 是多少？常用的量是多少？

A：(1)X 表示模板 DNA 量需要实验者在首次实验时进行摸索。首先对模板DNA 进行稀释（一般推荐 5-10 倍），然后模板量梯度上样，选择 CT 值落在 20-30 之间的最佳上样量。
(2)常用的量是逆转录 500-1000ng 总RNA，稀释 10 倍取 1μL cDNA 进行qPCR 实验。

Q：qPCR 实验结果的有效性？为什么建议Ct 值要大于 15？

A：a)有效性要满足三个条件：
（1）标准曲线：扩增效率范围:90-110%，对应斜率为 -3--3.5。R2＞0.98。 (扩增效率=10-1/斜率-1)，当斜率=-3.32 时，扩增效率=100%。
（2）扩增曲线：S 型曲线，且 Ct 值在 15－35 之间，阴性对照 Ct＞35 或无Ct 值。
（3）熔解曲线：为单一峰。

b)3-15 个循环的荧光值标准差的 10 倍是荧光阈值，Ct 值太小了会影响曲线。

Q：Rox 的作用？

A:ROX 是一种参比染料，其作用是标准化荧光定量反应中的非PCR 震荡，校正加样误差或者是孔与孔之间的误差，提供一个稳定的基线。

Q: 为什么扩增产物跑胶会有拖尾的现象？

A: 试剂中本身含有稳定性成分，本身不参与任何反应，稳定剂成分在电泳胶上面会表现出弥散状，不建议跑完后再电泳。

Q: 试剂比较粘稠，容易产生气泡，如何避免？

A: 可以先混合大体系，再分别加到离心管里面，最后加模板；

枪头加样不要把空气打进去，二档吸样，一档打样品；缓慢推出液体，不要吹打；沿着壁打入到孔内；另外配置完后，如果还有气泡，离心PCR管去除。如果是96孔板，移液完后轻轻敲96孔板。

Q：为什么从冰箱中拿出有淡黄色沉淀，还可以使用吗？

A：沉淀是稳定剂冻存后溶解度下降析出形成的，不影响产品性能。建议解冻后上下颠倒，枪头吹打混匀至沉淀消失。若仍未溶解，可离心后取液体，请避免取到该沉淀。

文档与文献

实时荧光定量PCR实验技术手册(反转定量) 查看文档

已发表文献

[1] Xia B, Shen X, He Y, et al. SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target. Cell Res. 2021;31(8):847-860. doi:10.1038/s41422-021-00519-4(IF:25.617)

[2] Wang S, Li Y, Zhong L, et al. Efficient gene editing through an intronic selection marker in cells. Cell Mol Life Sci. 2022;79(2):111. Published 2022 Jan 31. doi:10.1007/s00018-022-04152-1(IF:9.261)

[3] An LL, Zhao X, Gong XY, et al. Promoter Binding and Nuclear Retention Features of Zebrafish IRF Family Members in IFN Response. Front Immunol. 2022;13:861262. Published 2022 Apr 6. doi:10.3389/fimmu.2022.861262(IF:7.561)

[4] Zhao Y, Wang HP, Yu C, et al. Integration of physiological and metabolomic profiles to elucidate the regulatory mechanisms underlying the stimulatory effect of melatonin on astaxanthin and lipids coproduction in Haematococcus pluvialis under inductive stress conditions. Bioresour Technol. 2021;319:124150. doi:10.1016/j.biortech.2020.124150(IF:7.539)

[5] Shu C, Wang L, Zou C, et al. Function of Foxl2 and Dmrt1 proteins during gonadal differentiation in the olive flounder Paralichthys olivaceus [published online ahead of print, 2022 Jun 16]. Int J Biol Macromol. 2022;215:141-154. doi:10.1016/j.ijbiomac.2022.06.098(IF:6.953)

[6] Wang S, Huang J, Liu F, et al. Isosteviol Sodium Exerts Anti-Colitic Effects on BALB/c Mice with Dextran Sodium Sulfate-Induced Colitis Through Metabolic Reprogramming and Immune Response Modulation. J Inflamm Res. 2021;14:7107-7130. Published 2021 Dec 20. doi:10.2147/JIR.S344990(IF:6.922)

[7] Wang J, Hu R, Wang Z, et al. Establishment of Immortalized Yak Ruminal Epithelial Cell Lines by Lentivirus-Mediated SV40T and hTERT Gene Transduction. Oxid Med Cell Longev. 2022;2022:8128028. Published 2022 Mar 25. doi:10.1155/2022/8128028(IF:6.543)

[8] Liu W, Guan Y, Qiao S, et al. Antiaging Effects of Vicatia thibetica de Boiss Root Extract on Caenorhabditis elegans and Doxorubicin-Induced Premature Aging in Adult Mice. Oxid Med Cell Longev. 2021;2021:9942090. Published 2021 Aug 6. doi:10.1155/2021/9942090(IF:6.543)

[9] Qian Z, Liu C, Li H, et al. Osteocalcin Alleviates Lipopolysaccharide-Induced Acute Inflammation via Activation of GPR37 in Macrophages. Biomedicines. 2022;10(5):1006. Published 2022 Apr 27. doi:10.3390/biomedicines10051006(IF:6.081)

[10] Zhao X, Huang Y, Li X, et al. Full integration of nucleic acid extraction and detection into a centrifugal microfluidic chip employing chitosan-modified microspheres [published online ahead of print, 2022 Jun 27]. Talanta. 2022;250:123711. doi:10.1016/j.talanta.2022.123711(IF:6.057)

[11] Xu X, Wang H, Li X, Duan X, Wang Y. A novel ALG10/TGF-β positive regulatory loop contributes to the stemness of colorectal cancer. Aging (Albany NY). 2022;14(11):4858-4873. doi:10.18632/aging.204116(IF:5.955)

[12] Li F, Hu X, Wu J. Daidzein Activates Akt Pathway to Promote the Proliferation of Female Germline Stem Cells through Upregulating Clec11a [published online ahead of print, 2022 Jun 3]. Stem Cell Rev Rep. 2022;10.1007/s12015-022-10394-0. doi:10.1007/s12015-022-10394-0(IF:5.739)

[13] Ru M, Wang W, Zhai Z, et al. Nicotinamide mononucleotide supplementation protects the intestinal function in aging mice and D-galactose induced senescent cells. Food Funct. 2022;13(14):7507-7519. Published 2022 Jul 18. doi:10.1039/d2fo00525e(IF:5.396)

[14] Ye M, Gao R, Chen S, et al. Downregulation of MEG3 and upregulation of EZH2 cooperatively promote neuroblastoma progression. J Cell Mol Med. 2022;26(8):2377-2391. doi:10.1111/jcmm.17258(IF:5.310)

[15] Wang S, Wang X, Shao Y, et al. Synthesis and evaluation of 3-(phenylethynyl)-1,1'-biphenyl-2-carboxylate derivatives as new HIF-1 inhibitors. Bioorg Chem. 2021;116:105298. doi:10.1016/j.bioorg.2021.105298(IF:5.275)

[16] Guan X, Meng X, Zhu K, et al. MYSM1 induces apoptosis and sensitizes TNBC cells to cisplatin via RSK3-phospho-BAD pathway. Cell Death Discov. 2022;8(1):84. Published 2022 Feb 26. doi:10.1038/s41420-022-00881-1(IF:5.241)

[17] Liu H, Wang LL, Xu QH, et al. UHRF1 shapes both the trophoblast invasion and decidual macrophage differentiation in early pregnancy. FASEB J. 2022;36(4):e22247. doi:10.1096/fj.202101647RR(IF:5.192)

[18] Chen M, Wang J, Lin L, et al. Synergistic Regulation of Metabolism by Ca²⁺/Reactive Oxygen Species in Penicillium brevicompactum Improves Production of Mycophenolic Acid and Investigation of the Ca²⁺ Channel [published correction appears in ACS Synth Biol. 2022 Apr 15;11(4):1705]. ACS Synth Biol. 2022;11(1):273-285. doi:10.1021/acssynbio.1c00413(IF:5.110)

[19] Zhao X , Li X , Yang W , Peng J , Huang J , Mi S . An integrated microfluidic detection system for the automated and rapid diagnosis of high-risk human papillomavirus. Analyst. 2021;146(16):5102-5114. doi:10.1039/d1an00623a(IF:4.616)

[20] Chen H, Jiang Y, Liu R, et al. Curcumin Derivative C66 Suppresses Pancreatic Cancer Progression through the Inhibition of JNK-Mediated Inflammation. Molecules. 2022;27(10):3076. Published 2022 May 11. doi:10.3390/molecules27103076(IF:4.412)

[21] Su W, Qiu J, Mei Y, Zhang XE, He Y, Li F. A microfluidic cell chip for virus isolation via rapid screening for permissive cells [published online ahead of print, 2022 May 2]. Virol Sin. 2022;S1995-820X(22)00075-X. doi:10.1016/j.virs.2022.04.011(IF:4.327)

[22] Ding J, Mei S, Cheng W, Ni Z, Yu C. Curcumin treats endometriosis in mice by the HIF signaling pathway. Am J Transl Res. 2022;14(4):2184-2198. Published 2022 Apr 15. (IF:4.060)

[23] Zhang XY, Shen HH, Qin XY, et al. IL-27 promotes decidualization via the STAT3-ESR/PGR regulatory axis. J Reprod Immunol. 2022;151:103623. doi:10.1016/j.jri.2022.103623(IF:4.054)

[24] Zhu Q, Muyayalo KP, Xu QH, Wang J, Wang H, Liao AH. Prunella vulgaris can improve the pregnancy outcomes of experimental autoimmune thyroiditis rats by inhibiting Th1/Th17 immune responses. J Reprod Immunol. 2022;149:103469. doi:10.1016/j.jri.2021.103469(IF:4.054)

[25] Yang Q, Guo K, Zhou X, et al. Histopathology, antioxidant responses, transcriptome and gene expression analysis in triangle sail mussel Hyriopsis cumingii after bacterial infection. Dev Comp Immunol. 2021;124:104175. doi:10.1016/j.dci.2021.104175(IF:3.636)

[26] Liu Y, Wan HH, Tian DM, et al. Development and Characterization of High Efficacy Cell-Penetrating Peptide via Modulation of the Histidine and Arginine Ratio for Gene Therapy. Materials (Basel). 2021;14(16):4674. Published 2021 Aug 19. doi:10.3390/ma14164674(IF:3.623)

[27] Fu Y, Wang X, Zhang L, Ren Y, Hao L. Allograft inflammatory factor-1 enhances inflammation and oxidative stress via the NF-κB pathway in diabetic kidney disease. Biochem Biophys Res Commun. 2022;614:63-69. doi:10.1016/j.bbrc.2022.04.089(IF:3.575)

[28] Zhao Y, Wang XQ. VvMYB1 potentially affects VvTOR gene expression by regulating VvTOR promoter and participates in glucose accumulation. J Plant Physiol. 2022;272:153668. doi:10.1016/j.jplph.2022.153668(IF:3.549)

[29] Cheng Y, Zheng L, Yang C, Zhang W, Wang H. Propofol inhibits proliferation and migration of glioma cells by up-regulating lncRNA GAS5. Toxicol In Vitro. 2022;80:105321. doi:10.1016/j.tiv.2022.105321(IF:3.500)

[30] Yang B, Wang F, Zheng G. Transmembrane protein TMEM119 facilitates the stemness of breast cancer cells by activating Wnt/β-catenin pathway. Bioengineered. 2021;12(1):4856-4867. doi:10.1080/21655979.2021.1960464(IF:3.269)

[31] Yang X, Liu X, Song F, et al. Seasonal expressions of GPR41 and GPR43 in the colon of the wild ground squirrels (Spermophilus dauricus). Eur J Histochem. 2022;66(1):3351. Published 2022 Jan 21. doi:10.4081/ejh.2022.3351(IF:3.188)

[32] Tang Z, Yuan X, Bai Y, et al. Seasonal changes in the expression of PACAP, VPAC1, VPAC2, PAC1 and testicular activity in the testis of the muskrat (Ondatra zibethicus). Eur J Histochem. 2022;66(2):3398. Published 2022 May 2. doi:10.4081/ejh.2022.3398(IF:3.188)

[33] Zhu P, Chen Y, Wu F, Meng M, Ji K. Expression and promoter analysis of MEP pathway enzyme-encoding genes in Pinus massoniana Lamb. PeerJ. 2022;10:e13266. Published 2022 Apr 12. doi:10.7717/peerj.13266(IF:2.984)

[34] Zhai L, Chen W, Cui B, Yu B, Wang Y, Liu H. Overexpressed versican promoted cell multiplication, migration and invasion in gastric cancer. Tissue Cell. 2021;73:101611. doi:10.1016/j.tice.2021.101611(IF:2.466)

[35] Jiang J, Cai M. Cardamonin Inhibited IL-1β Induced Injury by Inhibition of NLRP3 Inflammasome via Activating Nrf2/NQO-1 Signaling Pathway in Chondrocyte. J Microbiol Biotechnol. 2021;31(6):794-802. doi:10.4014/jmb.2103.03057(IF:2.351)