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主要由 Tris、NaCl、EDTA 组成,所以简称为 TNE 缓冲液。该试剂主要用于吸收和荧光光谱学定量 RNA 和 DNA,只要考虑了污染物和缓冲液组分的作用,吸收测量时很直接简单的方法,荧光分析比A₂₆₀更不易受干扰。
操作流程:
1、用去离子水稀释 10×TNE 缓冲液(pH7.4)至 1×。
2、取1mL1×TNE 缓冲液吸入石英杯,放入单光束或双光束分光光度计中,在352nm处读值,仪器调零。该空白溶液作为双光束仪器的参照。对于单光束分光光度计,去除空白杯,插入含有DNA样品或标准品的石英杯,读数。在280nm(蛋白)、260nm(核酸)、230nm(肽、酚、尿素)重复该过程。
3 、用A₂₆₀读数代入方程计算核酸的浓度(C)。(公式较复杂,一般用第4点代替计算)
4、用A₂₆₀/A₂₈₀比值和A₂₃₀和A₃₂₅处的读值来估计核酸样品的纯度,比值在1.8~1.9显示 DNA 纯度高,比值在1.9~2.0显示 RNA 纯度高。
温馨提示:
为了您的自身安全,使用试剂前,请做好防护,如穿实验服,带手套等。
Product Description:
Composed mainly of Tris, NaCl, and EDTA, this reagent is therefore abbreviated as TNE Buffer. It is primarily used for the quantification of RNA and DNA via absorption and fluorescence spectroscopy. As long as the effects of contaminants and buffer components are considered, absorption measurement is a straightforward and simple method, while fluorescence analysis is less susceptible to interference than A₂₆₀(absorbance at 260 nm) measurement.
Operating Procedure:
1. Dilute the 10×TNE Buffer (pH 7.4) to 1× concentration using deionized water.
2. Pipette 1 mL of 1×TNE Buffer into a quartz cuvette, place it in a single-beam or double-beam spectrophotometer, and measure the absorbance at 352 nm to zero the instrument. This blank solution serves as a reference for double-beam instruments. For single-beam spectrophotometers, remove the blank cuvette, insert the quartz cuvette containing the DNA sample or standard, and record the absorbance. Repeat this process at 280 nm (for proteins), 260 nm (for nucleic acids), and 230 nm (for peptides, phenols, and urea).
3. Substitute the A₂₆₀(absorbance at 260 nm) reading into the equation to calculate the nucleic acid concentration (C). (The formula is relatively complex, so Step 4 is generally used instead for calculation.)
4. Estimate the purity of the nucleic acid sample using the A₂₆₀/A₂₈₀ratio and the absorbance values at 230 nm and 325 nm. A ratio of 1.8–1.9 indicates high-purity DNA, while a ratio of 1.9–2.0 indicates high-purity RNA.
Friendly Reminder:
For your personal safety, please take protective measures before using the reagent, such as wearing a lab coat and gloves.
| pH | pH 7.4 |
|---|
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | T1374038 | |
| 分析证书 | T1374038 |
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