TE

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m (TE buffer moved to TE: fit the other buffer names better)
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==TE buffer==
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==Purpose==
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10 mM [[Tris]]-HCl
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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.
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1 mM [[EDTA]]
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==Recipe==
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==10X pH 8.0 Recipe==
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{|border="1"
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*100mM Tris-Cl (pH 8.0)
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|+ '''10xTE'''
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*10mM EDTA (pH 8.0)
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!  for 1 liter from stock solutions
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|-
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|10 ml 1M Tris-HCl pH 7.5 or 8.0 (see notes) 
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|-
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|2 ml 0.5M EDTA pH 8.0
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|-
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|988 ml ddH<sub>2</sub>O
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|}
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{|border="1"
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|+ '''1xTE'''
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!  for 1 liter from stock solutions
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|-
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|1 ml 1M Tris-HCl pH 7.5 or 8.0 (see notes)  
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|-
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|0.2 ml 0.5M EDTA pH 8.0
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|-
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|988 ml ddH<sub>2</sub>O
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|}
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→ 1x TE is 10 mM [[Tris]]-HCl and 1 mM [[EDTA]]
==Notes==
==Notes==
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*Use Tris base and adjust the pH to 8.0 using HCl.
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*For the Tris-HCl use Tris base and adjust to desired pH using HCl.
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*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.
+
* 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.
[[Category:Material]] and [[Category:Buffers]]
[[Category:Material]] and [[Category:Buffers]]

Revision as of 10:19, 8 May 2008

Purpose

TE buffer is often used to store DNA and RNA. The EDTA in TE chelates Mg2+ and other divalent metals ions necessary for most causes of DNA and RNA degradation, suppressing these processes.

Recipe

10xTE
for 1 liter from stock solutions
10 ml 1M Tris-HCl pH 7.5 or 8.0 (see notes)
2 ml 0.5M EDTA pH 8.0
988 ml ddH2O


1xTE
for 1 liter from stock solutions
1 ml 1M Tris-HCl pH 7.5 or 8.0 (see notes)
0.2 ml 0.5M EDTA pH 8.0
988 ml ddH2O

→ 1x TE is 10 mM Tris-HCl and 1 mM EDTA

Notes

  • For the Tris-HCl use Tris base and adjust to desired pH using HCl.
  • TE buffer is often used to store DNA and RNA. The EDTA in TE chelates Mg2+ 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 Mg2+, 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 Mg2+ for subsequent reactions to proceed successfully. Each EDTA molecule chelates one Mg2+ ion.
  • 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. and
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