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Q:贴壁细胞消化后可不可以离心后加入Trizol呢?有影响吗?
可以,用少量PBs重悬,然后用trizol提取,不过提取要快,不能存放,因为细胞为活体,表达受外界环境影响,要是已经存放了两小时,建议重做。
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Q:胰腺组织RNA提取时在研磨时总是粘在研钵,怎么处理?有什么好的方法吗?
取出胰腺组织后立刻浸入液氮中,然后将其转移至研钵中(研钵要经180度干烤4-8小时,除去RNAase),持续研磨,一边研磨一边加液氮,研磨成粉末。此时要注意液氮的补充,如果组织化开就会出现组织粘在研钵内的情况。研磨成粉末后加入Trizol,此时,由于低温Trizol也会冻住,持续研磨至Trizol化开并澄清亮,就可以了。然后离心,进行下面的步骤。也可以使用匀浆器,很便宜,也很简单。当然要经去处RNAase处理。在冰上将组织研磨成匀浆即可。
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Q:What is the difference in the number of suspended cells and adherent cells?
Because of the difficulty of staining suspension cells, it is necessary to increase the number of cells and extend the time of culture. Staining of adherent cells is relatively easy, and if the number of cells is too large, sometimes the absorbance will exceed the reading of the microplate reader.
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Q:How many cells should be seeded in each well for a live cell test?
When standard 96-well plates were used, the minimum inoculum size of adherent cells was at least 1,000 cells per well (100 μl of medium) . The sensitivity for detecting white blood cells is relatively low, so a dose of no less than 2,500 cells per well (100 μl of medium) is recommended.
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Q:Can Cck-8 stain living cells?
No. Because the major component of CCK-8 is a water-soluble tetrazolium salt (WST-8) , and electrons in living cells are exchanged by the electron carrier PMS onto WST8 in the medium. CCK-8 can not stain cells because the resulting jazzans are also highly water-soluble.
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Q:How can the error due to CCK-8 residue on the gun head or hole wall be reduced?
CCK-8 reagent can be diluted with medium and mixed before adding sample.
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Q:Is it necessary to test whether the medium and CCK-8 react before the experiment?
If necessary, check it with a hole. Since there may be a redox substance in the medium, it is necessary to confirm that the medium reacts with CCK-8 before a formal experiment. The normal OD should be below 0.4.
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Q:CAN CCK-8 detect bacterial cells?
You can test for E. E. coli, but could not detect yeast cells. To 100 ΜL. Coli culture medium was added with 10 μl CCK-8 solution and cultured for 1-4 hours or overnight.
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Q:Storage conditions for CCK-8 reagent?
The storage condition of CCK-8 kit of Liji Biology: 4 ° C short-term storage, valid for 3 months; -20 ° C long-term storage, valid for 24 months. [ note ] : repeated freezing and thawing will result in higher background OD values.
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Q:CCK-8 test solution toxic to cells?
The CCK-8 solution itself is slightly toxic due to the presence of high concentrations of PMS. However, CCK-8 added to the medium was not toxic because it was diluted 10-fold. Therefore, prolonged incubation, such as overnight or for several days, is acceptable. The same cell culture medium after CCK -8 detection can also be used for other cell proliferation detection, such as crystal violet detection, neutral red detection or DNA fluorescence detection. Since each cell has a different tolerance to CCK-8, the viability of the cells after addition of CCK-8 in culture is first examined when a long period of culture is required.
"Heyuan Liji" is a joint venture between Heyuan Biotechnology (stock code: 688238) and Liji Biotechnology, specializing in the "Liji Biotechnology" and "Life-ilab" reagent brands