HotStart™ 2X Green qPCR Master Mix
Quantitative PCR (qPCR, also called Real-time PCR), is a popular technology for precise analysis of gene expression. It can be classified into two categories according to different methods, dye-based and probe-based, in which, dye-based method is more popular, convenient and less costly. Dye-based qPCR monitors real-time fluorescence of the dye binding to the double-stranded DNA to measure DNA amplification indirectly during each cycle. At a point where the fluorescence signal is distinctly detected over the background, Ct value (Cq value) can be determined. The obtained Ct values can be used to evaluate relative target abundance, or calculate absolute target quantities in reference to an appropriate standard curve. Our product HotStart™ 2X Green qPCR Master Mix provides superior specificity, robust amplification efficiency, ideal reproducibility and stability in quantifying target DNA or cDNA. It’s a 2X PreMix, taking advantage of a hot-start Taq polymerase combined with the antibody. Ideal Taq polymerase and suitable buffer will guarantee preferable specificity and high amplification speed. Green I in the mix which is a DNA intercalator will emit fluorescence when bounds to the double-stranded DNA amplified in each cycle and monitoring the fluorescence by the instrument allows for the indirect quantification of amplification products at real time. This reagent is supplied together with two ROX Reference Dye of different concentration which should be used to normalize the fluorescent signal intensity between reactions for instruments with the corresponding requirements. When conducting the experiment, use the SYBR® or SYBR/FAM scan mode setting on the real-time instrument.
- 1. Liang Quan, Yuan Xin, et al. "Peptide‐Enhanced Bioactive Hydrogel Combined with Photodynamic‐Phage Synergistic Antibacterial Therapy System Accelerates Infected Wound Healing." Adv Healthc Mater. 2025 Jul 8:e2500875 PMID: 40624966
- 2. Wei Wang, Yuankai Song, et al. "Overexpression of KRAS enhanced the stemness of esophageal cancer cells inhibited by overexpression of circ0043898." BMC Cancer. 2025 Jul 1;25:1039 PMID: 40597918
- 3. Yue Tian, Lei Wei, et al. "Magnetic mesoporous silica loaded with CGRP antagonist and curcumin coated with cerium oxide for the treatment of neuropathic pain." J Mater. Sci: Mater Eng. 20, 82 (19 June 2025)
- 4. Xiangying Ou, Juan Yang, et al. "Histone hyperacetylation disrupts spermatogonial stem cells homeostasis and impairs spermiogenesis." Stem Cell Res Ther. 2025 Jun 15;16:305 PMID: 40518506
- 5. Tianliang Ma, Qimeng Liu, et al. "Fused exosomal targeted therapy in periprosthetic osteolysis through regulation of bone metabolic homeostasis." Bioactive Materials Volume 50, August 2025, Pages 171-188
- 6. Jinrong Zhang, Hongming Huang, et al. "Erchen Decoction ameliorates the rat model of polycystic ovary syndrome by regulating the steroid biosynthesis pathway." Phytomedicine. 2025 Jul 25:143:156852 PMID: 40446578
- 7. Kimberly Paulsen, Rosenna Chan, et al. "KSHV miRNAs target STING to evade innate immunity and facilitate KSHV lytic reactivation from latency." Cell Rep. 2025 Jun 24;44(6):115741 PMID: 40413741
- 8. Qiqi Gao, Mengjia Zhang, et al. "Microsecond pulsed electric fields induce myocardial ablation by secondary mitochondrial damage and cell death mechanisms." Sci Rep 15, 10132 (2025)
- 9. Quan Lin, Shiwei Guan, Minghui Peng, Kailun Zhang, Hewei Zhang, Taoming Mo, Haibo Yu et al. "Comprehensive analysis of SQOR involvement in ferroptosis resistance of pancreatic ductal adenocarcinoma in hypoxic environments." Front Immunol. 2025 May 1:16:1513589. PMID: 40375994
- 10. Zhuzhen Shen, Jieli Zhang, et al. "Construction and Identification of Inflammation-Related TF–mRNA–miRNA Coexpression Network and Immune Infiltration in Parkinson's Disease." Parkinsons Dis. 2025 May 7:2025:2323585 PMID: 40370709
- 11. Libo He, Jinye Dang, et al. "Myriocin Restores Metabolic Homeostasis in dAGE-Exposed Mice via AMPK-PGC1α-Mediated Mitochondrial Activation and Systemic Lipid/Glucose Regulation." Nutrients. 2025 Apr 30;17(9):1549 PMID: 40362857
- 12. Miaoguan Peng, Yingying Zhang, et al. "Thiamine and METTL14 in Diabetes Management with Intensive Insulin Therapy." Biomedicines. 2025 Apr 17;13(4):980 PMID: 40299682
- 13. Elena Afanaseva, Antonio Barragan, et al. "TIMP1 secretion induced by Toxoplasma effector GRA24 via p38 MAPK signaling promotes non-disruptive parasite translocation across polarized brain endothelial monolayers." mSphere. 2025 May 27;10(5):e0010225 PMID: 40265926
- 14. Tianliang Ma, Qimeng Liu, et al. "Fused exosomal targeted therapy in periprosthetic osteolysis through regulation of bone metabolic homeostasis." Bioact Mater. 2025 Apr 8:50:171-188 PMID: 40248188
- 15. Zhixin Zhu, Lulu Jin, et al. "Inhalable Ce Nanozyme‐Backpacked Phage Aims at Ischemic Cerebral Injury by M1‐Microglia Hitchhiking." Adv Mater . 2025 Jul;37(27):e2419903 PMID: 40231579
- 16. Ielyzaveta Slarve, Yushan Wang, et al. "Isoform specific regulation of osteopontin by AKT2 in hepatocytes and livers." Cell Signal. 2025 Aug:132:111799 PMID: 40216173
- 17. Zheyu Zhang,Tianliang Ma,Qimeng Liu et al. "Exosomes Derived from Bone Marrow Mesenchymal Stem Cells Encapsulated in M2 Macrophage Cell Membrane Targeted to Inhibit Joint Periprosthetic Inflammation." ACS Appl Mater Interfaces. 2025 Apr 16;17(15):22279-22292 PMID: 40168527
- 18. Richard McKinnon Walsh, Joseph Ambrose, et al. "Depletion of tumor-derived CXCL5 improves T cell infiltration and anti-PD-1 therapy response in an obese model of pancreatic cancer." J Immunother Cancer. 2025 Mar 22;13(3):e010057. PMID: 40121029
- 19. Yanchen Zhang, Ende Wu, et al. "TSHR-targeting nucleic acid aptamer treats Graves’ ophthalmopathy via novel allosteric inhibition." bioRxiv. March 11, 2025.
- 20. Qi Chen, Hui He, et al. "Identification of Acanthopanax trifoliatus (L.) Merr as a Novel Potential Therapeutic Agent Against COVID-19 and Pharyngitis RSAD2: A pathogenic interferon-stimulated gene at the maternal-fetal interface of patients with systemic lupus erythematosus." Molecules. 2025 Feb 25;30(5):1055 PMID: 40076279
- 21. Guangzhao Yan, Yanyan Wang. "Mechanistic Study of palmatine in Regulating Pyroptosis in Sepsis Cells via Signaling Pathways." bioRxiv. February 20, 2025.
- 22. Chengzhe Lai, Jinghu Liu, et al. "The first-line antihypertensive nitrendipine potentiated the therapeutic effect of oxaliplatin by downregulating CACNA1D in colorectal cancer." Open Med (Wars). 2025 Feb 12;20(1):20241138 PMID: 39958978
- 23. Qimeng Liu, Tianliang Ma, et al. "Fused extracellular vesicles from M macrophages and humanumbilical cord mesenchymal stem cells for the targeted regulationof macrophage pyroptosis in periprosthetic osteolysis." J Extracell Vesicles.2024 Dec;13(12):e70028. PMID: 39711510
- 24. Michael Young, David M Booth, et al. "Transcriptional regulation in the absence of inositol trisphosphate receptor calcium signaling." Front Cell Dev Biol. 2024 Dec 6:12:1473210 PMID: 39712573
- 25. Mirza, Amar Naeem. "The Regulation of GLI1 in Basal Cell Carcinoma." Stanford University. 2021.
- 26. Fei Lou, Wenbin Zhou, et al. "VPS26 Moonlights as a β-Arrestin-like Adapter for a 7-Transmembrane RGS Protein in Arabidopsis thaliana." Biochemistry. 2024 Oct 28. PMID: 39467170
- 27. Jingwei Bi, Limin Zhang, et al. "Nanoarchitectonics of Injectable Biomimetic Conjugates for Cartilage Protection and Therapy Based on Degenerative Osteoarthritis Progression." Biomater Res. 2024 Sep10:28:0075. PMID: 39257895
- 28. Chunhai Luo, Haoran Xu, et al. "Meiotic chromatin-associated HSF5 is indispensable for pachynema progression and male fertility." Nucleic Acids Res. 2024 Aug 20:gkae701. PMID: 39162221
- 29. Ruo-Bing He, Wei Li, et al. "Aurantiamide mitigates acute kidney injury by suppressing renal necroptosis and inflammation via GRPR-dependent mechanism."Int Immunopharmacol. 2024 Sep 30:139:112745. PMID: 39059099
- 30. Austin T Gregg, Tianxi Wang, et al. "Botulinum neurotoxin serotype A inhibited ocular angiogenesis through modulating glial activation via SOCS3." Angiogenesis. 2024 Jun 26. PMID: 38922557
- 31. Min Zhu, Yunguan Wang, et al. "PKD1 mutant clones within cirrhotic livers inhibit steatohepatitis without promoting cancer."Cell Metab. 2024 Jun 13:S1550-4131(24)00191-8. PMID: 38901424
- 32. Yuanyuan Zou, Yakun Wang, et al. "Banxia Xiexin decoction combined with 5‐ASA protects against CPT‐11‐induced intestinal dysfunction in rats via inhibiting TLR4/NF‐κB signaling pathway." Immun Inflamm Dis. 2024 Jun;12(6):e1208. PMID: 38860759
- 33. Zheng Zhou, Xiyan Zheng, et al. "Mice hepatic organoids for modeling nonalcoholic fatty liver disease and drug response."Stem Cells Dev. 2024 Jun 26. PMID: 38814825
- 34. Raja Gopal Reddy Mooli, Bokai Zhu, et al. "Epigenetically active chromatin in neonatal iWAT reveals GABPα as a potential regulator of beige adipogenesis."Front Endocrinol (Lausanne). 2024 May 3:15:1385811. PMID: 38765953
- 35. Lijia Ni, Rui Shen, et al. "GlmS plays a key role in the virulence factor expression and biofilm formation ability of Staphylococcus aureus promoted by advanced glycation end products." Virulence. 2024 Dec;15(1):2352476. PMID: 38741276
- 36. Michael Young, David M. Booth, et al. "Transcriptional regulation in the absence of Inositol Trisphosphate Receptor Calcium Signaling."bioRxiv. April 20, 2024.
- 37. Robert Schauner, Jordan Cress, et al. "Single cell and bulk RNA expression analyses identify enhanced hexosamine biosynthetic pathway and O-GlcNAcylation in acute myeloid leukemia blasts and stem cells."Front Immunol. 2024 Mar 27:15:1327405. PMID: 38601153
- 38. Lin Wang, Ye Lu, et al. "Gegen Qinlian tablets delay Alzheimer's disease progression via inhibiting glial neuroinflammation and remodeling gut microbiota homeostasis."Phytomedicine. 2024 Jan 26:128:155394. PMID: 38569294
- 39. Daohong He, Huasheng Wu, et al. "Se/GSH-enriched Candida utilis ameliorates DSS-induced colitis in mice by preventing oxidative stress and improving immune responses." Food Bioscience Volume 58, April 2024, 103656.
- 40. Miaomiao Sun, Honghong Zhan, et al. "Dehydrocostus lactone alleviates irinotecan-induced intestinal mucositis by blocking TLR4/MD2 complex formation."Phytomedicine. 2024 Feb 3:128:155371. PMID: 38518649
- 41. Tianxi Wang, Satoshi Kaneko, et al. "SOCS3 regulates pathological retinal angiogenesis through modulating SPP1 expression in microglia and macrophages."Mol Ther. 2024 May 1;32(5):1425-1444. PMID: 38504518
- 42. Xudong Wang, Tianxi Wang, et al. "Photoreceptors inhibit pathological retinal angiogenesis through transcriptional regulation of Adam17 via c-Fos." Angiogenesis. 2024 Aug;27(3):379-395. PMID: 38483712
- 43. Arne L Ten Hoeve, Matias E Rodriguez, et al. "Hypermigration of macrophages through the concerted action of GRA effectors on NF-κB/p38 signaling and host chromatin accessibility potentiates dissemination." bioRxiv. 2024 Feb 6:2024.02.06.579146 PMID: 38370679
- 44. Jinyu Xu, Lianshuang Zhang, et al. "Ferritinophagy-mediated ferroptosis of spermatogonia is involved in busulfan-induced oligospermia in the mice."Chem Biol Interact. 2024 Feb 25:390:110870. PMID: 38220133
- 45. Hongbo Guo, Qiudi Li, et al. "Molecular determinants within the C-termini of L-HDAg that regulate hepatitis D virus replication and assembly." JHEP Rep. 2023 Nov 15;6(1):100961. PMID: 38192534
- 46. Hongbo Guo, Dan Liu, et al. "Drug repurposing screen identifies vidofludimus calcium and pyrazofurin as novel chemical entities for the development of hepatitis E interventions." Virol Sin. 2024 Feb;39(1):123-133. PMID: 37984761
- 47. Ronald Bronson, Junfang Lyu, et al. "Transcriptome Analysis Reveals Molecular Signature and Cell-type Difference ofHomo sapiensEndothelial-to-mesenchymal Transition." G3 (Bethesda). 2023 Dec 6;13(12):jkad243. PMID: 37857450
- 48. R McKinnon Walsh, Joseph Ambrose, et al. "Adipose-Tumor Crosstalk contributes to CXCL5 Mediated Immune Evasion in PDAC." bioRxiv. 2023 Aug 16;2023.08.15.553432. PMID: 37645755
- 49. Raja Gopal Reddy Mooli, Yerin Han, et al. "Hepatic ketogenesis regulates lipid homeostasis via ACSL1-mediated fatty acid partitioning." Res Sq. 2023 Jul 18:rs.3.rs-3147009
- 50. Peng Teng, Yuefeng Wu, et al. "Pulsed Field Ablation as a Precise Approach for Cardiac Arrhythmia Treatment via Cardiac Microenvironment Remodeling." Bioelectrochemistry. 2023 Jul 4;154:108502. PMID: 37453203
- 51. Xiaotian He, Yiyang Ma, et al. "SERPINB5 is a prognostic biomarker and promotes proliferation, metastasis and epithelial‐mesenchymal transition (EMT) in lung adenocarcinoma." Thorac Cancer. 2023 Jul 9. PMID: 37424293
- 52. Runjie Song, Peilan Guo, et al. "A novel polypeptide CAPG-171aa encoded by circCAPG plays a critical role in triple-negative breast cancer." Mol Cancer. 2023 Jul 5;22(1):104. PMID: 37408008
- 53. Xiaotian He, Dechang Zhao, et al. "Intrinsic Immunogenic Tumor Cell Death Subtypes Delineate Prognosis and Responsiveness to Immunotherapy in Lung Adenocarcinoma." Biology (Basel). 2023 Jun 2;12(6):808. PMID: 37372093
- 54. Jinsong Yang, Haibin Xu, et al. "Absolute quantification of viable Vibrio cholerae in seawater samples using multiplex droplet digital PCR combined with propidium monoazide." Front Microbiol. 2023 Jun 9:14:1149981. PMID: 37362935
- 55. Yanzhuo Yang, Qingting Yu, et al. "Palmitate lipotoxicity is closely associated with the fatty acid-albumin complexes in BV-2 microglia." PLoS One. 2023 Apr 20;18(4):e0281189. PMID: 37079515
- 56. Zhichao Tang, Shalakha Hegde, et al. "Chemical-guided SHAPE sequencing (cgSHAPE-seq) informs the binding site of RNA-degrading chimeras targeting SARS-CoV-2 5'untranslated region." bioRxiv. 2023 Oct 23:2023.04.03.535453. PMID: 37066172
- 57. Xiaotian He, Dechang Zhao, et al. "Intrinsic Immunogenic Tumor Cell Death Subtypes Delineate Prognosis and Responsiveness to Immunotherapy in Lung Adenocarcinoma." Biology 2023, 12(6), 808.
- 58. Junxing Zhao, Zhichao Tang, et al. "Cellular Target Deconvolution of Small Molecules Using a Selection-Based Genetic Screening Platform." ACS Cent Sci. 2022 Oct 26;8(10):1424-1434. PMID: 36313155
- 59. Appolinaire A Olou, Joe Ambrose, et al. "SHP2 regulates adipose maintenance and adipocyte-pancreatic cancer cell crosstalk via PDHA1." J Cell Commun Signal. 2022 Sep 8. PMID: 36074246
- 60. Yang Liu, Libo He, et al. "Pharmacological inhibition of sphingolipid synthesis reduces ferroptosis by stimulating the HIF-1 pathway." iScience. 2022 Jun 6;25(7):104533. PMID: 35784791
- 61. Ziming Li, Tengfei Zhao, et al. "A reactive oxygen species-responsive hydrogel encapsulated with bone marrow derived stem cells promotes repair and regeneration of spinal cord injury." Bioact Mater. 2022 May 9;19:550-568. PMID: 35600969
- 62. Yue Zhuo, Shujun Li, et al. "Targeting SNORA38B attenuates tumorigenesis and sensitizes immune checkpoint blockade in non-small cell lung cancer by remodeling the tumor microenvironment via regulation of GAB2/AKT/mTOR signaling pathway." J Immunother Cancer. 2022 May;10(5):e004113. PMID: 35577506
- 63. Yiting Wan, Yanhua Song, et al. "Upregulated Fibulin-1 Increased Endometrial Stromal Cell Viability and Migration by Repressing EFEMP1-Dependent Ferroptosis in Endometriosis." Biomed Res Int. 2022 Jan 28;2022:4809415. PMID: 35127942
- 64. Gao-Chao Han, Hai-Ming Jing, et al. "Effects of lanthanum nitrate on behavioral disorder, neuronal damage and gene expression in different developmental stages of Caenorhabditis elegans." Toxicology. 2022 Jan 15;465:153012. PMID: 34718030
- 65. Guo Z, Liu Y, et al. "Versatile biomimetic cantharidin-tellurium nanoparticles enhance photothermal therapy by inhibiting the heat shock response for combined tumor therapy." Acta Biomater. 2020;110:208-220. PMID: 32278084
Related Biological Data

Related Biological Data

Related Biological Data

Components | 1000 rxn with 10μL reaction 500 rxn with 20μL reaction 200 rxn with 50μL reaction |
5000 rxn with 10μL reaction 2500 rxn with 20μL reaction 1000 rxn with 50μL reaction |
10000 rxn with 10μL reaction 5000 rxn with 20μL reaction 2000 rxn with 50μL reaction |
HotStart™ 2X Green qPCR Master Mix | |||
50X ROX Reference Dye (low concentration) | |||
50X ROX Reference Dye (high concentration) | |||
Store all components at -20°C and protect from light. Avoid repeated freeze/thaw cycles as possible. |
Hot-start mechanism for optimized specificity
Superior sensitivity and accuracy
Highly reproducible Ct values over a broad dynamic range
Convenient 2X PreMix formats to simplify experimental steps