產(chǎn)品說明書
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產(chǎn)品簡介
TRIeasyTM Total RNA Extraction Reagent是一種適用于各種動植物、細菌組織、細胞的總RNA抽提試劑，具有極強的裂解能力，可在短時間內(nèi)裂解細胞和組織樣本，并有效抑制樣本中RNA的降解，保持RNA的完整性。樣品在該試劑中能夠充分裂解，之后加入氯仿離心分層，形成上清層、中間層和有機層(鮮紅色下層)，RNA分布在上層水相中，收集上清層后，經(jīng)異丙醇沉淀便可得到總RNA。提取的總RNA純度高，基本不含蛋白質(zhì)及基因組DNA，可直接用于Northern、點雜交、mRNA純化、體外翻譯、RT-PCR、poly(A)+選擇、RNA酶保護分析以及構建cDNA文庫等多種分子生物學實驗。
此外，樣品中的DNA和蛋白也能以沉淀的方式還原。乙醇沉淀能析出中間層的DNA，而在有機層中加入異丙醇能沉淀出蛋白。本品操作簡單快速，所有操作可在一小時內(nèi)完成。對少量的組織(50-100 mg)和細胞(5×106)以及大量的組織(≥1 g)和細胞(>107)均有較好的裂解效果。
產(chǎn)品信息
|
貨號 |
10606ES60 |
|
規(guī)格 |
100 mL |
儲存條件
2~8℃避光保存，有效期1年。
使用說明
表1 每毫升Total RNA Extraction Reagent能夠充分裂解的最大樣本量
|
樣本類型 |
樣本使用量 |
|
貼壁細胞 |
10 cm2培養(yǎng)面積 |
|
懸浮動植物細胞或酵母細胞 |
5×106-1×107個 |
|
細菌 |
107個 |
|
動物組織 |
50-100 mg |
|
植物組織(多糖和多酚含量不高的) |
50-100 mg |
【注】：過多的樣本量可能會導致裂解不充分，并使產(chǎn)物純度降低。
注意事項
1. 本產(chǎn)品中含有苯酚，具有毒性和腐蝕性。如果吸入體內(nèi)、接觸皮膚、吞食等會引起中毒、灼傷及其他身體傷害。使用時應穿戴防護物，如防護服裝、手套、眼罩、面罩等。如果不小心接觸到眼睛時，應立即用大量的水沖洗并前往醫(yī)院治療。
2. 請穿實驗服并佩戴一次性手套操作，避免RNase污染。
3. 需自備氯仿、異丙醇（新開封或提取RNA專用）、75%乙醇 (DEPC處理水配制)、DEPC和DEPC處理水。
4. 樣品用Total RNA Extraction Reagent勻漿后，如果不加入氯仿進行下游實驗，可先-70℃凍存，可保存一個月以上。
5. RNA沉淀在75%乙醇中，4℃可保存1周，-20℃可保存1年。
6. RNA半衰期比較短，易降解，建議抽提后盡快進行后續(xù)實驗。
7. 本產(chǎn)品僅作科研用途！
Ver.CN20230809
Q：10606和19201的區(qū)別是什么？
A：19201是濃縮形式，需要的試劑用量更少一點，提取的準確性和提取反應的速度更快一些。一般10606就可以滿足常規(guī)提取需求，如果客戶想好更好的提取效果和rnase抑制效果可以用19201。
Q: 用在貼壁細胞上總是不能完全從培養(yǎng)瓶（皿）脫落，是裂解不完全嗎？
A: 這并不意味著裂解不完全。此時，細胞膜實際已完全裂開，并釋放RNA，繼續(xù)后續(xù)實驗即可。
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