计算溶液所需的质量、体积或浓度。
BioReagent,无菌,适用于微生物学 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
TE/LiAc水溶液 (1.1×) 是一种制作酿酒酵母感受态的buffer。TE起缓冲液的作用,提供稳定的PH值环境;LiAc 可使酵母细胞处于一种短暂的感受状态,此时酵母能够摄取外源 DNA。使用阿拉丁的 TE醋酸锂混合液 (1.1×) 制作酵母感受态细胞,用 pGADT7 质粒检测转化效率可达 10⁴-10⁶ cfu/μg DNA 。
产品组分与配方:
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使用方法:
一,感受态制作方法
1、酵母菌种在 YPDA 平板上划线,挑新鲜的单菌落到3ml YPDA(用 50ml 离心管)中 30℃,250rpm 摇菌 12h,取 50ul,到 50ml YPDA (用250ml三角瓶)中,250rpm 摇菌到 OD600 约0.15-0.3,不要超过。
2、室温 700g,5min 离心收集菌体,弃上清,用 100ml YPDA 重悬,30℃,250rpm 摇菌到 OD600=0.4-0.5。
3、室温 700g,5min 离心收集菌体,弃上清,用 50ml 无菌 ddH₂O 重悬。
4、室温 700g,5min 离心收集菌体,弃上清,用 15ml 4℃ 预冷的 TE/LiAc Solution (1.1×) 。1500g,30s 离心,弃上清,用 0.6ml TE/liAC(1.1×) 重悬,此时可以用来转化酵母文库;也可以分装10 支感受态用来转化普通酵母质粒 (最好立即使用,也可放冰中保存不超过 3h)。
二,感受态转化方法
酵母转化实验操作对比表(文库质粒 vs 普通质粒)
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注意事项:
1、制作不同数量的感受态,按比例使用TE/LiAc 溶液 (1.1×),此种方法制作的酵母感受态不可 -80℃ 保存。
2、不开封的TE/LiAc 溶液 (1.1×)可在 4℃ 保存 2 年,开封后若发现染菌,停止使用。
3、本产品仅供科研使用,请勿用于医药、临床诊断或治疗,食品及化妆品等用途,请勿存放于普通住宅区。
4、本公司将不为任何不正常使用此产品时所发生的意外负责。
TE/LiAc Aqueous Solution (1.1×) is a buffer for preparing competent cells of Saccharomyces cerevisiae. TE functions as a buffer to maintain a stable pH environment; lithium acetate (LiAc) transiently renders yeast cells competent, enabling the uptake of exogenous DNA. Competent yeast cells prepared using TE/LiAc Solution (1.1×) from Aladdin exhibit a transformation efficiency of 10⁴-10⁶ cfu/μg DNA when assayed with pGADT7 plasmid.
Product Components and Formulation:
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Usage Protocol
I. Preparation of Competent Yeast Cells
1. Streak yeast strain on YPDA agar plate, pick a fresh single colony and inoculate into 3 mL YPDA medium in a 50 mL centrifuge tube. Incubate at 30 °C with shaking at 250 rpm for 12h. Transfer 50 μL of the overnight culture into 50 mL YPDA medium in a 250 mL Erlenmeyer flask, shake at 250 rpm until OD600 reaches approximately 0.15–0.3 (do not exceed this range).
2. Harvest cells by centrifugation at 700 g for 5 min at room temperature, discard supernatant, resuspend pellet in 100 mL YPDA medium, and shake at 30°C, 250 rpm until OD600 = 0.4–0.5.
3. Collect cells via centrifugation at 700 g for 5 min at room temperature, remove supernatant, and resuspend pellet in 50 mL sterile ddH₂O.
4. Centrifuge at 700 g for 5 min at room temperature, discard supernatant, and resuspend pellet in 15 mL pre-chilled (4°C) TE/LiAc Solution (1.1×). Centrifuge at 1500 g for 30 s and discard supernatant, then resuspend cells in 0.6 mL TE/LiAc Solution (1.1×).The competent cells can be used directly for yeast library transformation; alternatively, aliquot into 10 portions for routine yeast plasmid transformation. For optimal results, use immediately. If temporary storage is needed, keep on ice for no more than 3h.
II. Transformation Protocol for Competent Yeast Cells
Yeast Transformation Protocol – Library vs. Regular Plasmid (Comparative Table)
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Precautions
1. When preparing different quantities of competent cells, use 1.1× TE/LiAc Solution proportionally. Yeast competent cells prepared by this method cannot be stored at -80 °C.
2. Unopened 1.1× TE/LiAc Solution can be stored at 4 °C for 2 years. Discontinue use if microbial contamination is observed after opening.
3. This product is for research use only. It shall not be used for medical treatment, clinical diagnosis, food, cosmetics or other related applications. Do not store it in residential areas.
4. Our company shall not be liable for any accidents arising from improper use of this product.
通过匹配包装上的批号来查找并下载产品的 COA,每批产品都进行了严格的验证,您可放心使用!
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | T1524606 | |
| 分析证书 | T1524606 |