RN38-EASYspin Plus 植物RNA快速提取試劑盒

產(chǎn)品介紹：

     本公司推出EASYspin無苯酚、氯仿RNA快速提取技術(shù)基礎(chǔ)上，又成功研發(fā)基因組DNA清除柱技術(shù)可以有效清除gDNA殘留，得到的RNA一般不需要DNase消化，可用于反轉(zhuǎn)錄PCR、熒光定量PCR等實驗。獨特的裂解液迅速裂解細胞和滅活細胞RNA酶，植物RNA助提劑PLANTaid幫助結(jié)合多糖多酚并通過離心去除，然后裂解混合物用乙醇調(diào)節(jié)RNA結(jié)合吸附到基因組DNA清除柱，基因組DNA清除柱子同時吸附清除殘留的DNA, 然后RNA被選擇性洗脫濾過。濾過的RNA用乙醇調(diào)節(jié)結(jié)合條件后，RNA在高離序鹽狀態(tài)下選擇性吸附于離心柱內(nèi)硅基質(zhì)膜， 再通過一系列快速的漂洗－離心的步驟, 去蛋白液和漂洗液將細胞代謝物，蛋白等雜質(zhì)去除， 最后低鹽的RNase free H20將純凈RNA從硅基質(zhì)膜上洗脫。

產(chǎn)品特點：

1.完全不使用有毒的苯酚，氯仿等試劑，也不需要乙醇沉淀等步驟。 

2.簡捷，單個樣品操作一般可在25分鐘內(nèi)完成，世界上最簡單快速的試劑盒。 

3.獨有的植物RNA助提劑可以有效結(jié)合多糖多酚，提高清除效果。 

4.獨家研發(fā)成功基因組DNA清除柱技術(shù)可以有效清除gDNA殘留，得到的RNA一般不需要DNase消化，可用于反轉(zhuǎn)錄PCR、熒光定量PCR等實驗。 

5.適應(yīng)性極其廣泛，可以提取包括棉花、松針、冬青樹葉、葡萄葉片、等100多種國內(nèi)外試劑盒提取失敗的樣品。詳細樣品列表請參考公司主頁產(chǎn)品介紹。 

6.多次柱漂洗確保高純度，OD260/OD280典型的比值達1.9～2.2，基本無DNA殘留,可用于RT-PCR，Northern-blot和各種實驗。

使用RN38-EASYspin Plus 植物RNA快速提取試劑盒 發(fā)表文獻

1.     桃果實、花、根、葉：Isolation, characterisation and phylogenetic analysis of resistance gene    analogues in a wild species of peach (Prunus kansuensis).Canadian Journal of Plant Science,    2011, 91(6): 961-970

2.     櫻桃花、葉、顎等各部位：Over-expression of the PaAP1 gene from sweet cherry (Prunus avium    L.)    causes early floweri.Journal of Plant Physiology, 2012,Available online 1 December    2012

3.     洋蔥根、莖、蕾、葉、雌雄蕊等各部位：Cloning and Expression Analysis of A Putative B Class MADS-box    Gene of AcPI in Onion. Scientia Agricultura Sinica, 2012, 45(23):4759-4769

4.     蕪菁：Isolation and Functional Characterisation of the Genes Encoding Δ8-Sphingolipid       Desaturase from Brassica rapa. Journal of Genetics and Genomics Volume 39, Issue 1, January    2012,    Pages 47–59

5.     蕪  菁 1 ：EXPRESSION, DIVERGENCE AND EVOLUTION OF THE CALEOSIN GENE FAMILY IN BRASSICA RAPA. Arch. Biol. Sci., Belgrade, 65 (3), 863-876, 2013 DOI:10.2298/ABS1303863H

6.     番茄葉：Effect of Low Temperature Stress on the Expression of ProDH Gene and the Activities of the Proline Dehydrogenase in Leaves of Tomato Seedling. Chinese Agricultural Science  Bulletin 2012,28(10):132-135

7.     梔子葉：Isolation of High Quality Total RNA from Gardenia jasminoides Eills.Chinese Agricultural    Science Bulletin.2012, 28(27):194-198

8.     油桐果實：Cui Qinqin, Han Xiaojiao, Chen Yicun, Zhan Zhiyong, Lin Liyuan, Wang Yangdong.     Isolation and Expression Characteristics of Biotin Carboxyl Carrier Protein Coding Gene（VfBCCP)  from Vernicia fordii.SCIENTIA SILVAE SINICAE. 2012, 48(8): Available online August

9.     油桐果實1：Selection of Reliable Reference Genes for Gene Expression Studies Using Real-Time    PCR in Tung Tree during Seed Development. PLoS ONE, 2012, 7(8): e43084

10. 紫菜：Molecular cloning and expression analysis of ribosomal protein S7 gene from Porphyra    haitanensis. JOURNAL OF FISHERIES OF CHINA, 2011, 35（12）：1814-1821

11. 石斛：Molecular characterization of a mitogen-activated protein kinase gene DoMPK1 in Dendrobium officinale. Acta Pharmaceutica Sinica, 2012, 47 (12): 1703-1709

12. 石斛1：ESTs Analysis Reveals Putative Genes Involved in Symbiotic Seed Germination in Dendrobium officinale. Symbiotic Germination Genes in D. officinale. August 2013 | Volume 8 | Issue 8 | e72705

13. 大豆：RNA-seq Analysis Reveals Ethylene-Mediated Reproductive Organ Development and Abscission in Soybean(Glycine max L. Merr.). Plant Mol Biol Rep, 2012, published online: 4 Dec, 2012

14. 大豆1：Construction of ethylene regulatory network based on the phytohormones related gene transcriptome profiling and prediction of transcription factor activities in soybean. Acta Physiol Plant, 2012, published online: 12 Dec, 2012

15. 紅花玉蘭：Expression Analysis of MAwuAG in Different Organs and Developmental Stages of Magnolia wufengensis. Chinese Bulletin of Botany, 2013, 48 (2): 1–5

16. 毛桃：Cloning and Phylogeny Analysis of PpAP2 Floral Homologous Genes in Peach. Chinese Agricultural Science Bulletin, 2013, 29(7): 99-104

17. 五倍子：Cloning and characterisation of a phenylalanine ammonia-lyase gene from Rhus chinensis. Plant Cell Rep, 2013, published online：15 March, 2013

18. :五倍子1：Cloning, characterization and expression of chalcone synthase from medicinal plant Rhus chinensis.J. Plant Biochem. Biotechnol. DOI 10.1007/s13562-013-0231-9

19. 青杄 ：cDNA Cloning and Bioinformatic Analysis of the sPPa1 Gene form Picea wilsonii. Plant Science Journal, 2012, 30(40): 394-401

20. 青杄 1：cDNA Cloning and Bioinformatic Analysis of PsbO Gene from Picea wilsonii.Life Science Research, 2012, 16(3): 201-206

21. 青杄 2：Cloning and Tissue Expression Analysis of PwPSAF in Picea wilsonii. SCIENTIA SILVAE SINICAE. Vol. 49，No. 10, Oct. 2013.

22. 洋蔥：Molecular Cloning and Transcriptional Analysis of the Putative AGAMOUS Homolog AcAG in Onion (Allium cepa. Plant Mol Biol Rep, DOI 10.1007/s11105-013-0607-y

23. 木瓜：XsFAD2 gene encodes the enzyme responsible for the high linoleic acid content in oil accumulated in Xanthoceras sorbifolia seeds. JOURNAL ARTICLE. 2013-6-17.

24. 木瓜1：Two novel diacylglycerol acyltransferase genes from Xanthoceras 2 sorbifolia are responsible for its seed oil content. GENE-38688; No. of pages: 9; 4C:

25. 柑橘：Efficient auto-excision of a selectable marker gene from transgenic citrus by combining the Cre/loxP system and ipt selection. Plant Cell Rep, DOI 10.1007/s00299-013-1470-x

26. 柑橘1：Expression Analysis of Three Phloem-specific Promoters in Transgenic Poncirus trifoliata. Acta Horticulturae Sinica. 2014, 41(1): 1–8.

27. 柑橘2： Activation of three pathogen-inducible promoters in transgenic citrus (Citrus sinensis Osbeck) after Xanthomonas axonopodis pv. citri infection and wounding. Plant Cell Tiss Organ Cult. DOI 10.1007/s11240-013-0423-y.

28. 茶梅花瓣：Comparison and Analysis of Methods of Extracting Total RNA from Petals of Camellia sasanqua. Chinese Agricultural Science Bulletin.2013,29(28):129-133.

29. 梔子：Isolation of High Quality Total RNA fromGardenia jasminoides Eills. Chinese Agricultural Science Bulletin. 2012, 28(27):194-198

30. 丹參：Genome-wide analysis and molecular dissection of the SPL gene family in Salvia miltiorrhiza. 2014 Jan;56(1):38-50. doi: 10.1111/jipb.12111. Epub 2013 Nov 20.

31. 牡丹：Transcriptome Comparison Reveals Key Candidate Genes Responsible for the Unusual Reblooming Trait in Tree Peonies. Genes Responsible for Reblooming in Tree Peonies. November 2013 | Volume 8 | Issue 11 | e79996

32. 東南景天：Role of sulfur assimilation pathway in cadmium hyperaccumulation by Sedum alfredii Hance. Ecotoxicology and Environmental Safety. Volume 100, February 2014, Pages 159–165.

33. 山蒼子：Identification of appropriate reference genes for normalizing transcript expression by quantitative real?time PCR in Litsea cubeba. TECHNICAL NOTE. Mol Genet Genomics (2013) 288:727–737, DOI 10.1007/s00438-013-0785-1

34. 木本植物：Heterologous gene silencing induced by tobacco rattle virus (TRV) is efficient for pursuing functional genomics studies in woody plants. ORIGINAL PAPER. Plant Cell Tiss Organ Cult, DOI 10.1007/s11240-013-0393-0

35. 棉花：Analysis of sea-island cotton and upland cotton in response to Verticillium dahliae infection by RNA sequencing. Sun et al. BMC Genomics 2013, 14:852 /1471-2164/14/852.

36. 桃子：Biochemical changes and defence responses during the development of peach gummosis caused by Lasiodiplodia theobromae. Eur J Plant Pathol (2014) 138:195–207, DOI 10.1007/s10658-013-0322-4.

37. 桃子1：Carbohydrate metabolism changes in Prunus persica gummosis infected with Lasiodiplodia theobromae. Phytopathology "First Look" paper ? http://dx.doi.org/10.1094/PHYTO-01-13-0025-R ? posted 11/27/2013.

38. 海棠：The Malus crabapple transcription factor McMYB10 regulatesanthocyanin biosynthesis during petal coloration. Scientia Horticulturae 166 (2014) 42–49.

39. 海藻：A rapid and sensitive method for field detection of Prorocentrum donghaiense using reverse transcription-coupled loop-mediated isothermal amplification. Harmful Algae 29 (2013) 31–39.

40. 油茶：Establish a cDNA-AFLP Technology System in Camellia oleifera. Molecular Plant Breeding, 2013, Vol.11, No.5, 611-616.

41. 亞洲百合：Transcriptomic analysis of Asiatic lily in the process of vernalization via RNA-seq. Mol Biol Rep. DOI 10.1007/s11033-014-3250-2.

42. 毛泡桐：Dynamic expression of novel and conserved microRNAs and their targets in diploid and tetraploid of Paulownia tomentosa. Biochimie xxx (2014) 1e10.

43. 人參：Cloning and Sequence Analysis Squalene Epoxidase Gene in Panax gin-seng. Journal of Jilin Agricultural University 2014, 36(2): 149-152,17

44. 雪蓮：Cloning and Sequence Analysis of rbcs Gene from Sasussured involucrdta Kar. et Kir. Chinese Agricultural Science Bulletin 2014, 30(15): 261-267

45. 柑橘3：Secreted Expression of Cecropin B Gene Enhances Resistance to Xanthomonas axonopodis pv. citri in Transgenic Citrus sinensis‘Tarocco’ Acta Horticulturae Sinica 2014, 41(3): 417–428 http: // www. ahs. ac. cn

46. 菊花：Stem apex detoxification culture markedly improved severalphysiological characters of chrysanthemum ‘YUTAI’. Plant Cell Tiss Organ Cult 2014, DOI 10.1007/s11240-014-0541-1

47. 蕎麥和擬南芥：Ectopic expression of FaesAP3, a Fagopyrum esculentum (Polygonaceae) AP3 orthologous gene rescues stamen development in an Arabidopsis ap3 mutant. Gene 2014, 550(2): 200–206

48. 油松：Differential expression of SLOW WALKER2 homologue in ovules of female sterile mutant and fertile clone of Pinus tabulaeformis. Russian Journal of Developmental Biology 2014, 45(2): 78-84

49. 玫瑰花：Precise spatio-temporal modulation of ACC synthase by MPK6 cascade mediates the response of rose flowers to rehydration. The Plant Journal 2014, 79(6): 941–950

50. 棉花和擬南芥：Functional characterization of GhAKT1, a novel Shaker-like K+ channel gene involved in K+ uptake from cotton (Gossypium hirsutum). Gene 2014, 545(1): 61–71

51. 棉花和擬南芥1：Upland Cotton Gene GhFPF1 Confers Promotion of Flowering Time and Shade-Avoidance Responses in Arabidopsis thaliana. PLoS ONE 2014, 9(3): e91869. doi:10.1371/journal.pone.0091869

52. 白楊：Poplar GATA transcription factor PdGNC is capable of regulating chloroplast ultrastructure, photosynthesis, and vegetative growth in Arabidopsis under varying nitrogen levels. Plant Cell Tiss Organ Cult 2014, DOI 10.1007/s11240-014-0536-y

53. 毛果楊：Molecular characterization of the SPL gene family in Populus trichocarpa. BMC Plant Biology 2014, 14: 131

54. 葛根：Molecular cloning and characterization of an isoflavone 7-O-glucosyltransferase from Pueraria lobata. Plant Cell Reports 2014, 33(7), 1173–1185

55. 百合：Cloning and Expression Analysis of Actin Gene（lilyActin）from Lily. Acta Horticulturae Sinica 2013, 40(7): 1318–1326

56. 百合1： Vernalization of Oriental hybrid lily ‘Sorbonne’: changes in physiology metabolic activity and molecular mechanism.Molecular Biology Reports 2014, DOI 10.1007/s11033-014-3545-3

57. 黃鵪菜：Transcriptome Sequencing and De Novo Analysis of Youngia japonica Using the Illumina Platform. PLoS ONE 2014, 9(3): e90636. doi:10.1371/journal.pone.0090636

58. 棉花1：Gibberellin Overproduction Promotes Sucrose Synthase Expression and Secondary Cell Wall Deposition in Cotton Fibers. PLoS ONE 2014, 9(5): e96537. doi:10.1371/journal.pone.0096537

59. 蘋果：Low Medium pH Value Enhances Anthocyanin Accumulation in Malus Crabapple Leaves. PLoS ONE 2014, 9(6): e97904. doi:10.1371/journal.pone.0097904

其它公司多糖多酚植物RNA提取試劑盒失敗原因和解決方案

很多植物RNA的樣品由于含有大量的多糖、多酚、代謝產(chǎn)物、色素等成分，造成RNA提取過程中氧化、褐化、降解、由于植物品種的多樣性造成情況更加復(fù)雜。手工的CTAB類的方法提取因為時間太長，太繁瑣，手工方法不在討論之列。一直以來沒有一款好的試劑盒包括qiagen、promega等進口試劑盒也無法滿足科研工作者對植物RNA提取的要求。

下面我們來分析一下植物RNA為什么不能提取成功的原因：

市面上最常見的RNA提取試劑盒無非是兩種：種：TRIzol改良類方法（包括溶液型的和離心柱型的）、第二種：直接裂解過柱子的方法（離心柱）

種試劑盒失敗的原因1：RNA市面上面最流行的方法就是Trizol，或者Triol改良，或者Trizol加離心柱一類的改良方法。trizol也就是異硫氰酸胍/苯酚/氯仿原理一步法的方法最適合的對象是動物源性的組織細胞，針對普通多糖多酚低的植物性的材料，TRIzol類原理產(chǎn)品也可以提取。但是多糖、多酚、次級代謝產(chǎn)物豐富的情況下，trizol類方法無法防止多糖多酚對于RNA/DNA分相的干擾，要么殘留大量DNA，要么殘留大量多糖、多酚或者次級代謝產(chǎn)物，氧化破壞RNA，或者殘留這些多糖多酚，色素代謝產(chǎn)物等抑制下游的反轉(zhuǎn)錄等反應(yīng)。限于技術(shù)水平的限制，市面上絕大多數(shù)的國產(chǎn)廠家是使用trizol的方法進行改良，無論是不是加了離心柱。但是實踐證明，改良不能從根本上解決問題。判斷是否試劑盒使用這種改良的方法非常簡單：是否裂解液含有苯酚的味道和使用氯仿，如果使用到了氯仿就是TRIzol方法的改良。

第二種試劑盒失敗的原因：直接裂解過柱子的方法是目前最先進的方法，但是也是技術(shù)含量最高的方法。這個方法采用裂解液（不含苯酚，氯仿）直接裂解，RNA/DNA同時過柱子，然后在柱子上面直接分離RNA/DNA，所以，這種方法的優(yōu)點在于，避免了使用trizol在多糖多酚下不能成功分離RNA/DNA的弊端、第二在于，不使用有毒的苯酚氯仿。但是正是因為其技術(shù)先進，所以難度很高，國內(nèi)廠家包括進口公司有兩個技術(shù)難點一直沒有突破。，裂解液的成分必須針對去除多糖多酚進行研發(fā)添加去多糖多酚，代謝產(chǎn)物成分。否則會同樣碰到多糖多酚干擾提取的問題。第二、和trizol原理不同，直接過柱法DNA/RNA同時加到吸附柱上去。如何去除DNA是個難點。否則會殘留大量DNA。兩個技術(shù)難點的沒有掌握導(dǎo)致了國內(nèi)公司包括的第二種試劑盒失敗。國外公司因為沒有掌握難點，裂解液里面沒有去除多糖多酚成分，所以包括qiagen的盒子也常常不能成功提取植物RNA樣品。

部分成功樣品：

植物：棉花、海棠、黑加侖、煙草、擬南芥、虎杖、大豆、草莓、冬青、月季花雌蕊、薔薇、沙棘、冬棗、蘆薈、仙人掌、報春花、水稻、玉米、唐菖蒲、櫻桃、白玉蘭、毛白楊、櫻花、葡萄、百合花、百合葉子雌蕊雄蕊、紫菜、綠藻、香蕉、水仙花、青花菜、地被菊、蘋果、梅花、番茄、石斛、毛桃、苧麻、慈姑、葛根、甘肅桃、玫瑰花、檳榔果、甜糖菊、硅藻、牡丹、胡楊、油桐果、梨子皮、板栗花序、青皮云杉、紅樹根、鐵線蕨、黃瓜、小麥葉子種子、番木瓜、甘薯、紫薯、油松、油茶、馬尾松、蕪菁、毛果楊、木薯、大葉落地生根、山杏、旱柳、桉樹、琵琶花果、丹參、人參、西洋參、梔子、洋蔥、紅豆杉、梨樹葉、五倍子、泡桐、西瓜、芍藥、雪蓮等等，其中包括qiagen無法提取的黑加侖、冬青、月季、松針、葡萄葉片等，promega無法提取的海棠等樣品、均可用該產(chǎn)品成功提取。