ToolsQuant II Fast RT Kit

ToolsQuant II Fast RT Kit

   ToolsQuant II Fast RT Kit is designed for the first strand cDNA synthesis with high efficiency and able to rapidly remove genomic DNA (gDNA). To avoid the contamination of gDNA, the kit contains gDNA Eraser that effectively remove gDNA (42°C, 3 min). It takes only 15 min (42°C) to complete the synthesis of first strand cDNA by using the innovative RT enzyme. This product also has a high affinity for RNA, which enables efficient and sensitive reverse transcription of any template, such as GC rich and complicated secondary structures of template RNA, leading to high yields of cDNA.

產品型號 規格 單價 庫存 購買數量
KRT-BA06-2 100 rxns. - -
KRT-BA06 25 rxns. - -
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產品特點
  1. High efficiency:  Reverse transcription efficiency above 95%.

  2. gDNA Eraser included:remove genomic DNA in 3min

  3. High performance:Tolerate to GC-rich template

  4. Fast: Only 21 min experimental procedure

  5. Error rate: ~1.0 x10-3

操作流程

       

實驗數據

TOOLSQuant II Fast RT Competition Test:

參考文獻
  1. Wen YC, Lin YW, Chu CY, Yang YC, Yang SF, Liu YF, Hsiao M, Lee WJ, Chien MH. Melatonin-triggered post-transcriptional and post-translational modifications of ADAMTS1 coordinately retard tumorigenesis and metastasis of renal cell carcinoma. J Pineal Res. 2020 Sep;69(2):e12668. 

  2. Yang WB, Hsu CC, Hsu TI, Liou JP, Chang KY, Chen PY, Liu JJ, Yang ST, Wang JY, Yeh SH, Chen RM, Chang WC, Chuang JY. Increased activation of HDAC1/2/6 and Sp1 underlies therapeutic resistance and tumor growth in glioblastoma. Neuro Oncol. 2020 Oct 14;22(10):1439-1451. 

  3. Huang CS, Ho JY, Chiang JH, Yu CP, Yu DS. Exosome-Derived LINC00960 and LINC02470 Promote the Epithelial-Mesenchymal Transition and Aggressiveness of Bladder Cancer Cells. Cells. 2020 Jun 7;9(6):1419. 

  4. Fang CH, Lin YT, Liang CM, Liang SM. A novel c-Kit/phospho-prohibitin axis enhances ovarian cancer stemness and chemoresistance via Notch3-PBX1 and β-catenin-ABCG2 signaling. J Biomed Sci. 2020 Mar 13;27(1):42. 

  5. Lang YD, Chen HY, Ho CM, Shih JH, Hsu EC, Shen R, Lee YC, Chen JW, Wu CY, Yeh HW, Chen RH, Jou YS. PSPC1-interchanged interactions with PTK6 and β-catenin synergize oncogenic subcellular translocations and tumor progression. Nat Commun. 2019 Dec 16;10(1):5716. 

  6. Li CH, Chang YC, Hsiao M, Liang SM. FOXD1 and Gal-3 Form a Positive Regulatory Loop to Regulate Lung Cancer Aggressiveness. Cancers (Basel). 2019 Nov 28;11(12):1897. 

  7. Huang HC, Chao CC, Wu PH, Chung HY, Lee HY, Suen CS, Hwang MJ, Cai BH, Kannagi R. Epigenetic silencing of the synthesis of immunosuppressive Siglec ligand glycans by NF-κB/EZH2/YY1 axis in early-stage colon cancers. Biochim Biophys Acta Gene Regul Mech. 2019 Feb;1862(2):173-183. 

  8. Hsu RM, Zhong CY, Wang CL, Liao WC, Yang C, Lin SY, Lin JW, Cheng HY, Li PY, Yu CJ. Golgi tethering factor golgin-97 suppresses breast cancer cell invasiveness by modulating NF-κB activity. Cell Commun Signal. 2018 Apr 27;16(1):19. 

  9. Chang CJ, Lu CC, Lin CS, Martel J, Ko YF, Ojcius DM, Wu TR, Tsai YH, Yeh TS, Lu JJ, Lai HC, Young JD. Antrodia cinnamomea reduces obesity and modulates the gut microbiota in high-fat diet-fed mice. Int J Obes (Lond). 2018 Feb;42(2):231-243. 

  10. Wu YY, Hou BH, Lee WC, Lu SH, Yang CJ, Vaucheret H, Chen HM. DCL2- and RDR6-dependent transitive silencing of SMXL4 and SMXL5 in Arabidopsis dcl4 mutants causes defective phloem transport and carbohydrate over-accumulation. Plant J. 2017 Jun;90(6):1064-1078. 

  11. Chang CJ, Lin CS, Lu CC, Martel J, Ko YF, Ojcius DM, Tseng SF, Wu TR, Chen YY, Young JD, Lai HC. Ganoderma lucidum reduces obesity in mice by modulating the composition of the gut microbiota. Nat Commun. 2015 Jun 23;6:7489. Erratum in: Nat Commun. 2017 Jul 11;8:16130. 

  12. Hou CY, Lee WC, Chou HC, Chen AP, Chou SJ, Chen HM. Global Analysis of Truncated RNA Ends Reveals New Insights into Ribosome Stalling in Plants. Plant Cell. 2016 Oct;28(10):2398-2416. 

  13. Yang ZS, Huang SW, Wang WH, Lin CY, Wang CF, Urbina AN, Thitithanyanont A, Tseng SP, Lu PL, Chen YH, Wang SF. Identification of Important N-Linked Glycosylation Sites in the Hemagglutinin Protein and Their Functional Impact on DC-SIGN Mediated Avian Influenza H5N1 Infection. Int J Mol Sci. 2021 Jan 13;22(2):743.

  14. Chu CY, Lee YC, Hsieh CH, Yeh CT, Chao TY, Chen PH, Lin IH, Hsieh TH, Shih JW, Cheng CH, Chang CC, Lin PS, Huang YL, Chen TM, Yen Y, Ann DK, Kung HJ. Genome-wide CRISPR/Cas9 knockout screening uncovers a novel inflammatory pathway critical for resistance to arginine-deprivation therapy. Theranostics. 2021 Jan 25;11(8):3624-3641.

  15. Wu YS, Ho JY, Yu CP, Cho CJ, Wu CL, Huang CS, Gao HW, Yu DS. Ellagic Acid Resensitizes Gemcitabine-Resistant Bladder Cancer Cells by Inhibiting Epithelial-Mesenchymal Transition and Gemcitabine Transporters. Cancers (Basel). 2021 Apr 22;13(9):2032.

常見問題

Q1.利用Quant II合成的cDNA進行qPCR後,結果相較於以前跑的數值CT值變高,是否正常?

A:最佳的RT反應RNA劑量為1ug,可以調整濃度後看看,並且請再確認Control和目標Gene的比值是否有差異

 

Q2.合成的cDNA效果不好,應該如何改善?

A:請再次確認目標基因的長度,Quant II最多合成長度為4kb。

 

Q3.Quant II RT反應,最後95C的步驟若沒有做,是否會有什麼影響?

A:為了將Enzyme inactive,建議要完成95C(3分鐘)的步驟。

 

Q4.Quant II的Primer成分及濃度是什麼?

A:包含Random primer及Oligo dT primer,濃度為1:1

 

Q5.Quant II可否用於Ecoli系統的RNA?

A:可以, Quant II所附的Primer mix是含有Random Primer,因此可以用於Ecoli系統

 

Q6.Quant II的input量低於50ng可以嗎?

A:建議不要,依protocol最低的建議量為50ng,除非受限樣本的情況下再進行嘗試

 

Q7.Quant II是否可以作流感病毒的RT?

A:可以, 有randon primer即可

 

Q8.若客戶使用Quant II時,希望僅使用Oligo dT或是random primer的情況下,Primer要使用多少濃度?

A:建議參考protocol說明,在final體積20ul的情況下,Oligo dT及Random Primer濃度為50pmol,則Specific Primer為5pmol

 

Q9.Quant II是否可以自行調整Buffer,以及不做gDNA去除的步驟?

A:若要調整不用去除gDNA的步驟請客戶自行嘗試,但Buffer的量還是請依照Datasheet中提到的gDNA removal reaction及RT reaction的比例調整

 

Q10.Qunat II中的Buffer是否含有Rnase Inhibitor?

A:有的

 

Q11.Mastermix裡面是否含有RNaseH?

A:無 

 

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