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TE: Difference between revisions
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==Notes== | ==Notes== | ||
*Use Tris base and adjust the pH to 8.0 using HCl. | *Use Tris base and adjust the pH to 8.0 using HCl. | ||
*TE buffer is often used to store DNA and RNA. The [[EDTA]] in TE chelates Mg<sup>2+</sup> and other divalent metals ions necessary for most causes of DNA and RNA degradation, suppressing these processes. However, downstream reactions like restriction digests, PCR, ligations, and reverse transcription typically require Mg<sup>2+</sup>, potentially making the presence of EDTA in the reaction problematic. So, when using DNA or RNA that was suspended in TE, you should keep track of the amount of EDTA in the mix to make sure there is still enough Mg<sup>2+</sup> for subsequent | *TE buffer is often used to store DNA and RNA. The [[EDTA]] in TE chelates Mg<sup>2+</sup> and other divalent metals ions necessary for most causes of DNA and RNA degradation, suppressing these processes. However, downstream reactions like restriction digests, PCR, ligations, and reverse transcription typically require Mg<sup>2+</sup>, potentially making the presence of EDTA in the reaction problematic. So, when using DNA or RNA that was suspended in TE, you should keep track of the amount of EDTA in the mix to make sure there is still enough Mg<sup>2+</sup> for subsequent reactions to proceed successfully. Each EDTA molecule chelates one Mg<sup>2+</sup> ion. | ||
*Some protocols use TE 10:0.1 with 0.1 mM EDTA to reduce the interaction of the EDTA with downstream applications. | *Some protocols use TE 10:0.1 with 0.1 mM EDTA to reduce the interaction of the EDTA with downstream applications. | ||
*Some people use TE buffers with different pH's for different applications. For example, DNA is stored at pH 8 to reduce depurination, which is acid catalyzed, while RNA is stored at a slightly lower pH (7.5) because degradation of RNA is base-catalyzed. Most downstream reactions will not be influenced by the slightly different pH storage conditions. | *Some people use TE buffers with different pH's for different applications. For example, DNA is stored at pH 8 to reduce depurination, which is acid catalyzed, while RNA is stored at a slightly lower pH (7.5) because degradation of RNA is base-catalyzed. Most downstream reactions will not be influenced by the slightly different pH storage conditions. | ||
