Tris–HCl,Tris(Hydroxymethyl) Aminomethane,Tris Hydroxymethyl Aminomethane Buffer

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Ricipes of buffers for RNA experiments with TRIS 2019-04-12 17:44:36

Tris(Hydroxymethyl) Aminomethane (TRIS) buffer solution is a biological reagent, which is a buffer solution commonly used in biological preparation and medical preparation. It is commonly used in: gel electrophoresis buffer, such as TAE or TBE; preparation of Laemmli buffer; preparing running buffer and loading buffer together with glycine and SDS; binding buffer and an eluent in anion exchange chromatography; RNA hybridization and DNA cleavage. Below we will introduce you the ricipes of buffers for RNA experiments with TRIS.

1. RNA Ligase Buffer

(10×)

Reagent	Quantity (for 1.0 mL)	Final concentration (10×)
Tris–HCl (1 M, pH 8.0)	500 µL	0.5 M
MgCl₂(1 M)	150 µL	0.15 M
Dithiothreitol (DTT) (0.5 M)	66 µL	33 mM
H₂O	284 µL

(2×)

Reagent	Quantity (for 1.0 mL)	Final concentration (2×)
ATP (100 mM)	10 µL	1 mM
Tris–HCl (1 M, pH 8.0)	200 µL	200 mM
MgCl₂ (1 M)	20 µL	20 mM
DTT (1 M)	6.6 µL	6.6 mM
DMSO	200 µL	20% (v/v)
Glycerol (ultrapure)	300 µL	30% (v/v)
H₂O	to 1 mL	-

2. RNA isolation buffer

Tris-HCl (10 mM, pH 7.6)

EDTA (1 mM)

Sodium dodecyl sulfate (SDS) (0.5%, w/v)

Prepare from DEPC-treated H2O and RNase-free stocks of Tris-HCl, EDTA, and SDS.

3. RNA Binding

Washing Buffer:

1× Tris-buffered saline (TBS)

0.05% Tween 20

2 mM MgCl2 (RNase-free)

Blocking Buffer:

10 mM Tris (pH 7.5, RNase-free)

137 mM NaCl (RNase-free)

3 mM KCl (RNase-free)

2 mM MgCl2 (RNase-free)

2% Bovine serum albumin (BSA)

10 µg/mL ssDNA

10 µg/mL yeast tRNA

Combine all ingredients except ssDNA and yeast tRNA.

4. Lysis Buffer for Yeast RNA

Reagent	Quantity (for 100 mL)	Final concentration
Tris–HCl (1 M, pH 6.8)	5 mL	50 mM
NaCl (1 M)	10 mL	100 mM
EDTA (0.5 M, pH 8.0)	2 mL	10 mM
ddH₂O	to 100 mL	–

5. RNA Annealing Buffer (10x)

0.5 M NaCl

100 mM dithiothreitol

100 mM MgCl2

100 mM Tris-HCl (pH 7.5)

6. RNA transcription buffer

Reagent	Amount to add	Final concentration
MgCl₂ (1 M)	300 μL	30 mM
NaCl (5 M)	100 μL	50 mM
Spermidine (1 M)	100 μL	10 mM
Tris-HCl (1 M) (pH 7.9)	2 mL	200 mM
H₂O, nuclease-free	7.6 mL

7. Transcription Buffer for RNA Polymerase (5×)

Reagent	Quantity (for 10 mL)	Final concentration
Tris–HCl (1 M, pH 8.0)	2 mL	200 mM
MgCl₂ (1 M)	400 µL	40 mM
NaCl (5 M)	0.5 mL	250 mM
Spermidine (100 mM)	1 mL	10 mM
H₂O	6.1 mL	–

In the use of TRIS, it should be noted that the pH of Tris buffer is greatly affected by temperature and concentration, so the environment should be considered during the preparation process.

Edited by Suzhou Yacoo Science Co., Ltd.

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