Trizol (总RNA抽提试剂)
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产品编号 R0016
数量
¥ 589.00
产品包装:
100ml
产品简介
使用说明
说明书下载
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产品问答

碧云天生产的Trizol是一种用于细胞或组织总RNA抽提的试剂。本产品采用和Invitrogen公司的TRIzol完全相似的原理和方法,抽提的方法和步骤完全相同。
Trizol的颜色和TRIzol相同,加入氯仿后上层呈无色,下层呈紫红色,便于吸取上层水相。
Trizol对动植物细胞或组织及细菌的总RNA抽提均适用。
Trizol可以抽提长达15 kb的RNA,也可以抽提microRNA等小RNA。抽提小RNA时宜-70℃沉淀过夜。
Trizol抽提所得RNA无DNA和蛋白污染。一般所得RNA溶于DEPC水后的A260/280值为1.8-2.0。
裂解细胞或和组织共匀浆时,Trizol可以保持样品中RNA的完整性,即可以有效抑制RNA的降解。
每一百万细胞用Trizol抽提可得5-15μg RNA;每毫克组织用Trizol抽提可得1-10μg RNA。产量因细胞和组织不同而异。
抽提两个样品约需一小时。
Trizol抽提所得RNA可直接用于Northern,点杂交,纯化mRNA,体外翻译,RNase protection assay,cDNA克隆,以及RT-PCR;也可以用于基因表达芯片分析、高通量测序(deep sequencing)等对RNA质量要求较高的情况。
碧云天生产的Beyozol(R0011)Trizol(R0016)的成分有细微差别,实际抽提效果无任何显著差异。
每100ml Trizol可以抽提100个六孔板中的样品或100个50-80mg的组织样品。
包装清单:

产品编号 产品名称 包装
R0016 Trizol 100ml
说明书 1份

保存条件:
4℃保存,一年有效。
注意事项: 
需自备氯仿,异丙醇,DEPC,75%乙醇(DEPC水配制),和DEPC水。DEPC(ST036)DEPC水(R0021)可向碧云天订购。
所有离心管,枪头及相关溶液都必须无RNA酶污染。耐高温器物可150℃烘烤4小时以去除RNA酶,其它器物去除RNA酶可考虑用0.01%的DEPC水浸泡过夜,然后灭菌,烘干。溶液需用DEPC水配制。加0.01%(体积比) diethylpyrocarbonate(DEPC)至重蒸水或Milli-Q级水中,处理过夜,灭菌即成DEPC水。
使用冻存的细胞或组织抽提总RNA的效果通常比新鲜的细胞或组织差一些。因为在细胞或组织冻融过程中一些细胞或组织内的RNase会被释放出来并剪切样品。如果不能及时抽提RNA,推荐先加入适量Trizol,并裂解样品后冻存。
必须戴一次性手套操作,且尽量不要对着RNA样品呼气或说话,以防RNA酶污染。建议戴一次性口罩操作。
Trizol含有毒物质苯酚,避免接触皮肤或吸入。为防止溅入眼睛,请戴防护眼镜或使用透明保护屏。如皮肤接触Trizol,请立即用大量去垢剂和水冲洗,如仍有不适,请听取医生意见。
Trizol抽提总RNA的同时,理论上也可抽提蛋白和DNA,但未经测试。有兴趣者可按Invitrogen公司的TRIzol的操作步骤进行操作。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1. 细胞裂解或组织匀浆。
a. 贴壁细胞
吸尽培养液,每10平方厘米细胞加入1ml Trizol。一般六孔板每孔加1ml Trizol,12孔板每孔加0.5ml Trizol。晃动3-5下,再用枪吹打2-3下,确保全部裂解,然后吸至离心管中。
b. 悬浮细胞
离心收集细胞,吸尽液体,每五百万至一千万动植物或酵母细胞,或一千万细菌,加入1ml Trizol。用枪吹打或适当vortex,确保全部裂解。某些酵母和细菌如裂解不充分,可用匀浆器匀浆,确保全部裂解。
c. 组织
先将组织剪切成小块,放入普通玻璃匀浆器内。每50mg-80mg组织加入1ml Trizol,匀浆。对于RNA完整性要求比较高的情况,推荐先液氮冷冻组织块,然后在低温下用研钵研碎组织,随后再加入Trizol进行总RNA抽提。
2. 对于某些蛋白,多糖或脂含量很高的细胞或组织,Trizol裂解后可能会有不溶物或油脂状漂浮物。需12,000g 4℃离心10分钟,然后吸取澄清的Trizol裂解产物至一新的离心管中。
3. 室温放置5分钟,使样品充分裂解。
4. 每毫升 Trizol加入0.2ml氯仿,vortex混匀或猛烈晃动15秒,室温放置2-3分钟。
5. 12,000g 4℃离心15分钟,然后吸取含总RNA的上层无色水相至一新的离心管中,每毫升Trizol约可吸取0.5-0.55ml。
6. 按每毫升最初的Trizol加入0.5ml异丙醇,颠倒数次混匀,室温沉淀10分钟。如果希望提取microRNA等小RNA,推荐-70℃沉淀过夜。
7. 12,000g 4℃离心10分钟,在管底可见RNA沉淀,弃上清。
8. 每毫升最初的Trizol加入1ml 75%乙醇(DEPC水配制),vortex或颠倒混匀,
9. 7,500g 4℃离心5分钟,弃上清。再用离心机甩一下(>5,000rpm, 离心1秒),小心吸尽液体。
10. 待RNA略干后,加入20μl DEPC水溶解,-70℃冻存。注意:切勿让RNA过分干燥,否则将极难溶解,且测出的A260/280值会低于1.6。
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