mRNA建库试剂盒，含全套mRNA抓取以及mRNA建库试剂盒。
Hieff NGS® Ultima Dual-mode mRNA Library Prep Kit是兼容Illumina和MGI双平台的mRNA转录组文库构建试剂盒，本试剂盒将cDNA二链合成与Endprep、dA-tailing进行合并，极大地缩减建库时间。二链合成模块配有两种Buffer，客户可根据需要进行常规建库或链特异性建库。本产品适用于起始模板为10 ng-4 μg不同来源真核生物总RNA样本。经过mRNA分离、片段化、双链cDNA合成、末端修复、加A尾、接头连接和文库扩增，总RNA样品最终转化为兼容Illumina®和MGI®平台测序的文库。
试剂盒包含两个独立模块，BOX-I的核心为纯化mRNA所需的oligo (dT)磁珠。BOX-II包含mRNA片段化试剂，反转录试剂，常规和链特异性ds-cDNA合成，以及后续建库所需的所有试剂。其中链特异性二链合成Buffer中将dTTP替换为dUTP，使cDNA第二链中掺入dUTP，而本试剂盒使用的高保真DNA聚合酶无法扩增含尿嘧啶的DNA模板，实现链特异性。提供的所有试剂都经过严格的质量控制和功能验证，最大程度上保证了文库构建的稳定性和重复性。
- 建库简便性：二链合成与末端修复，加A流程一步完成。
- 片段更集中，文库均一性好：不同片段条件对应不同大小文库，文库测序均一性好，无5’或3’偏好性。
- 投入量范围广：兼容10 ng ~ 4 μg的Total RNA投入量。
- 链特异性及非链特异性兼容：优化的扩增酶使用不同的buffer可简便得到链特异或者非链特异文库。
- 测序平台兼容：Illumina和MGI双测序平台兼容，搭配不同测序平台接头，即可完成对应平台的测序文库。
产品应用：mRNA建库
1、温和可控的mRNA片段化条件
表 mRNA片段化条件推荐
- 根据目的文库大小选择合适的片段化程序
- 根据样本质量调整片段化程序
图. mRNA不同打断时间对应的双链cDNA片段范围。以Universal Human Reference RNA(Agilent: 740000)为例。
2、不同样本文库峰型一致性较好
选用人、小鼠、小麦和拟南芥，使用同一投入量以及相同的实验条件进行mRNA建库，对比样本的文库产量以及文库峰型，最终结果显示，不同样本浓度均符合要求，且文库峰型一致性较高。
3、文库均一性较好
分析文库测序数据，看文库5端到3端的覆盖度，结果显示，建库试剂盒无偏好性，文库覆盖度很均一。
Box I：2-8°C保存。
Box II：-25~-15℃保存。
[1]Tian S, Zhang B, He Y, et al. CRISPR-iPAS: a novel dCAS13-based method for alternative polyadenylation interference. Nucleic Acids Res. 2022;50(5):e26. doi:10.1093/nar/gkac108(IF:19.160)
[2]Lin Z, Luo P, Lin X, et al. Effects of a sulfated glycosaminoglycan from Sepia esculenta ink on transcriptional and metabolic profiles of Saccharomyces cerevisiae. Carbohydr Polym. 2022;276:118715. doi:10.1016/j.carbpol.2021.118715(IF:10.723)
[3]Zhang Y, Wu T, Li F, et al. FGF19 Is Coamplified With CCND1 to Promote Proliferation in Lung Squamous Cell Carcinoma and Their Combined Inhibition Shows Improved Efficacy. Front Oncol. 2022;12:846744. Published 2022 Apr 7. doi:10.3389/fonc.2022.846744(IF:5.738)
[4]Yang J, Cui Y, Zhang X, et al. Maize PPR278 Functions in Mitochondrial RNA Splicing and Editing. Int J Mol Sci. 2022;23(6):3035. Published 2022 Mar 11. doi:10.3390/ijms23063035(IF:6.208)
[5]Hu P, Guo S, Yang S, et al. Stachytine Hydrochloride Improves Cardiac Function in Mice with ISO-Induced Heart Failure by Inhibiting the α-1,6-Fucosylation on N-Glycosylation of β1AR. Front Pharmacol. 2022;12:834192. Published 2022 Feb 8. doi:10.3389/fphar.2021.834192(IF:5.988)
[6]Lin Z, Luo P, Huang D, Wu Y, Li F, Liu H. Multi-omics based strategy for toxicity analysis of acrylamide in Saccharomyces cerevisiae model. Chem Biol Interact. 2021;349:109682. doi:10.1016/j.cbi.2021.109682(IF:5.168)
[7]Wu Z, Li H, Zhang Y, et al. Liver transcriptome analyses of acute poisoning and recovery of male ICR mice exposed to the mushroom toxin α-amanitin. Arch Toxicol. 2022;96(6):1751-1766. doi:10.1007/s00204-022-03278-2(IF:6.168)
[8]Liu S, Liu J, Wang Y, et al. Differentially expressed genes induced by β-caryophyllene in a rat model of cerebral ischemia-reperfusion injury. Life Sci. 2021;273:119293. doi:10.1016/j.lfs.2021.119293(IF:6.780)
[9]Zhao H, Tang X, Wu M, et al. Transcriptome Characterization of Short Distance Transport Stress in Beef Cattle Blood. Front Genet. 2021;12:616388. Published 2021 Feb 10. doi:10.3389/fgene.2021.616388(IF:4.772)
[10]Sun X, Lv W, Wang Y, et al. Mrgprb2 gene plays a role in the anaphylactoid reactions induced by Houttuynia cordata injection. J Ethnopharmacol. 2022;289:115053. doi:10.1016/j.jep.2022.115053(IF:5.195)
[11]Liu T, Zhang Y, Cao Y, et al. Optimization of oncolytic effect of Newcastle disease virus Clone30 by selecting sensitive tumor host and constructing more oncolytic viruses. Gene Ther. 2021;28(12):697-717. doi:10.1038/s41434-020-0145-9(IF:4.184)
[12]Zhong Y, Zhao W, Tang Z, et al. Comparative transcriptomic analysis of the different developmental stages of ovary in red swamp crayfish Procambarus clarkii. BMC Genomics. 2021;22(1):199. Published 2021 Mar 21. doi:10.1186/s12864-021-07537-x(IF:4.547)
[13]Liu Y, Han R, Zhou L, et al. Comparative performance of the GenoLab M and NovaSeq 6000 sequencing platforms for transcriptome and LncRNA analysis [published correction appears in BMC Genomics. 2022 Jan 26;23(1):81]. BMC Genomics. 2021;22(1):829. Published 2021 Nov 17. doi:10.1186/s12864-021-08150-8(IF:4.547)
[14]Zheng J, Tian J, Wang S, et al. Stachydrine hydrochloride suppresses phenylephrine-induced pathological cardiac hypertrophy by inhibiting the calcineurin/nuclear factor of activated T-cell signalling pathway. Eur J Pharmacol. 2020;883:173386. doi:10.1016/j.ejphar.2020.173386(IF:5.195)
[15]Yangchun G, Yufeng W, Dainan C, et al. Transcriptome analysis reveals decreased immunity under heat stress in Mauremys mutica. Aquaculture,Volume 531,2021,https://doi.org/10.1016/j.aquaculture .2020.735894(IF:5.135)
[16]Diao G, Huang J, Zheng X, et al. Prostaglandin E2 serves a dual role in regulating the migration of dendritic cells. Int J Mol Med. 2021;47(1):207-218. doi:10.3892/ijmm.2020.4801(IF:5.314)
[17]Huang X, Liang C, Yang H, et al. Curcumin induces apoptosis and inhibits the growth of adrenocortical carcinoma: Identification of potential candidate genes and pathways by transcriptome analysis. Oncol Lett. 2021;21(6):476. doi:10.3892/ol.2021.12737(IF:3.111)
[18]Wen X, Liu S, Cui M. Effect of BRCA1 on the Concurrent Chemoradiotherapy Resistance of Cervical Squamous Cell Carcinoma Based on Transcriptome Sequencing Analysis. Biomed Res Int. 2020;2020:3598417. Published 2020 Jun 21. doi:10.1155/2020/3598417(IF:3.246)
[19]Ding CY, Ma YM, Li B, et al. Identification and Functional Analysis of Differentially Expressed Genes in Myzus persicae (Hemiptera: Aphididae) in Response to Trans-anethole. J Insect Sci. 2022;22(1):3. doi:10.1093/jisesa/ieab094(IF:2.066)
[20]Qian D, Tian J, Wang S, et al. Trans-cinnamaldehyde protects against phenylephrine-induced cardiomyocyte hypertrophy through the CaMKII/ERK pathway. BMC Complement Med Ther. 2022;22(1):115. Published 2022 Apr 25. doi:10.1186/s12906-022-03594-1(IF:2.838)
[21]Liu QH, Wang ZY, Tang JW, et al. Comparative transcriptome analysis of diurnal alterations of liver glycogen structure: A pilot study. Carbohydr Polym. 2022;295:119710. doi:10.1016/j.carbpol.2022.119710(IF:10.723)
[22]Wei W, Li Y, Wang C, et al. Diterpenoid Vinigrol specifically activates ATF4/DDIT3-mediated PERK arm of unfolded protein response to drive non-apoptotic death of breast cancer cells. Pharmacol Res. 2022;182:106285. doi:10.1016/j.phrs.2022.106285(IF:10.334)
[23]Zhang S, Deng L, Zhao L, Wu C. Genome-wide binding analysis of transcription factor Rice Indeterminate 1 reveals a complex network controlling rice floral transition. J Integr Plant Biol. 2022;64(9):1690-1705. doi:10.1111/jipb.13325(IF:9.106)
[24]Zhao H, Wu M, Tang X, et al. RNA-seq Based Transcriptome Analysis Reveals The Cross-Talk of Macrophage and Adipocyte of Chicken Subcutaneous Adipose Tissue during The Embryonic and Post-Hatch Period. Front Immunol. 2022;13:889439. Published 2022 Jul 15. doi:10.3389/fimmu.2022.889439(IF:8.786)
[25]Yang W, Wang K., Xia S, et al. Coordinating High Yield and Superior Eating Quality of Rice: A Case Study of Hybrid Varieties Derived from Longke638S and Jing4155S. Agronomy 2022, 12, 1628. https://doi.org/10.3390/agronomy12071628(IF:3.949)
[26]Zhou X, Liu N, Jiang X, et al. A chromosome-scale genome assembly of Quercus gilva: Insights into the evolution of Quercus section Cyclobalanopsis (Fagaceae). Front Plant Sci. 2022;13:1012277. Published 2022 Sep 23. doi:10.3389/fpls.2022.1012277(IF:6.627)
