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Buffer formulation for RNA experiments using TRIS

Release time:

2021-09-14

Tris(hydroxymethyl)aminomethane (TRIS) buffer is a biological reagent, which is commonly used in biological preparations and pharmaceutical preparations. Commonly used: gel electrophoresis buffer, such as TAE or TBE; preparation of Laemmli buffer; preparation of running buffer and loading buffer together with glycine and SDS; binding buffer and eluent in anion exchange chromatography; RNA hybridization And DNA cutting. Below we will introduce to you the buffer formulations for TRIS RNA experiments.

1. RNA ligase buffer (10×)

Reagent Quantity (1.0 mL) Final Concentration (10×)

Tris-HCl (1M, pH 8.0) 500 μl 0.5 medium

Magnesium chloride 2 (1 M) 150 μl 0.15 M

Dithiothreitol (DTT) (0.5 M) 66 µL 33 mm

H 2 O 284 μl

2. RNA ligase buffer (2×)

Reagent Quantity (1 .0 mL) Final concentration (2×)

ATP (100 mm) 10 μl 1 mm

Tris-HCl (1 M, pH 8.0) 200 μl 200 mm

Magnesium chloride 2 (1 M) 20 μl 20 mm

DTT (1 M) 6.6 μl 6.6 mm

DMSO 200 μl 20% (v/v)

Glycerin (ultra pure) 300 μl 30% (v/v)

H 2 O to 1 ml ——

3. Yeast RNA Lysis Buffer

Reagent Quantity (100ml) Final Concentration

Tris-HCl (1 M, pH 6.8) 5 ml 50 mm

Sodium chloride (1 M) 10 ml 100 mm

EDTA (0.5 M, pH 8.0) 2 ml 10 mm

ddH 2 0 to 100 ml ——

4. RNA transcription buffer

Reagent Add amount Final concentration

Magnesium chloride 2 (1 M) 300 μl 30 mm

Sodium chloride (5 M) 100 μl 50 mm

Spermidine (1 M) 100 μl 10 mm

Tris- H Cl (1 M) (pH 7.9) 2 ml 200 mm

H 2 O, nuclease-free 7.6 ml

5. RNA polymerase transcription buffer (5×)

Reagent Quantity (10ml) Final Concentration

Tris-HCl (1 M, pH 8.0) 2 ml 200 mm

Magnesium chloride 2 (1 M) 400 μl 40 mm

Sodium chloride (5 M) 0.5 ml 250 mm

Spermidine (100 mM) 1 ml 10 mm

H 2 O 6.1 ml ——

6. RNA binding washing buffer:

1× Tris Buffered Saline (TBS)

0.05% Tween 20

2 mM MgCl2 (without RNase)

Blocking buffer:

10 mM Tris (pH 7.5, without RNase)

137 mM NaCl (without RNase)

3 mM KCl (without RNase)

2 mM MgCl2 (without RNase)

2% Bovine Serum Albumin (BSA)

10 µg/mL single-stranded DNA

10 µg/mL yeast tRNA

Combines all ingredients except ssDNA and yeast tRNA.

When using TRIS, it should be noted that the pH value of Tris buffer is greatly affected by temperature and concentration. Therefore, the environment must be considered during the preparation process. Desun's professional TRIS manufacturer has reached a long-term cooperative relationship with many manufacturers. , Welcome to order and consult.