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Takara

serochem PEI轉(zhuǎn)染試劑配制說明書

時間:2023/6/6閱讀:369
分享:
serochem PEI轉(zhuǎn)染試劑是一種高性能轉(zhuǎn)染試劑,設(shè)計用于穩(wěn)健、低成本和可擴展的瞬時基因表達(dá)。PEI是HEK293和CHO懸浮培養(yǎng)物瞬時轉(zhuǎn)染zui常用的試劑。下面以PEI Prime為例,來說明serochem PEI轉(zhuǎn)染試劑配制。

一、實驗材料:

serochem PEI轉(zhuǎn)染試劑100 mg(Prime-P100-100MG)、100 mL細(xì)胞培養(yǎng)級水、10 mL 0.1 M USP級氫氧化NA、(0.1或0.2)μm注射器過濾器、刻度100毫升容器、無菌等分容器(聚丙烯或聚乙|烯)

二、實驗設(shè)備:

分析天平、pH計、生物安全柜或潔凈室外殼、過濾設(shè)備(如注射器、真空泵)

三、配制流程:

1. 準(zhǔn)確稱量100 mg PEI Prime,將粉末溶于80mL細(xì)胞培養(yǎng)級水中。攪拌幾分鐘來溶解粉末。

注意:混合后可能會殘留微量固體。如果存在,將在稍后的步驟中移除,這些不會影響性能。

2. 逐漸向PEI Prime中加入0.1 M氫氧化NA溶液,同時用pH計監(jiān)測,將pH調(diào)節(jié)至7.0 +/- 0.1,在加入氫氧化NA之后充分混合。注意:使用少于10ml的氫氧化NA溶液

3. 添加細(xì)胞培養(yǎng)級水,使PEI Prime溶液使總體積達(dá)到100ml。

4. 在無菌環(huán)境中,如清潔的生物安全機柜或潔凈室,對溶液進(jìn)行無菌過濾,用(0.1或0.2)μm注射器過濾器將溶液無菌過濾到無菌儲存容器中。

注意:建議使用PES過濾膜;尼龍膜也可行。

5. 在4°C的溫度下給容器貼上標(biāo)簽,可儲存長達(dá)6個月。

注意:不要冷凍,不要使用冷凍過一次以上的等分試樣。


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