EMSA+说明书英文版 Non-Radioactive EMSA User Manual VER.3.11 This user manual is suitable for the production SIDET001, SIDET003, SIDET004.Revised 3/2007
2. Kit components (1)The kits* of EMSA-Basic include the follows: 10X Binding Buffer (-20 o C) 1 Vial Poly (dI: dC) (dI: dC) (-20 o C) 1 Vial 6X Loading Buffer (4 o C) 1 Vial Cold oligonucleotides (-20 o C) 1 Vial Biotin-Labeled Probe (-20 o C) 1 Vial Streptavidin...
组成成分• 10X Binding Buffer, 1 mL • Biotin-EBNA Control DNA, 50 µL • Unlabeled EBNA DNA, 50 µL • EBNA Extract, 125 µL • Poly (dI·dC), 125 µL • 50% Glycerol, 500 µL • 1% NP-40, 500 µL • 1 M KCl, 1 mL • 100mM MgCl2, 500 µL...
probe is 50-100 times that of the biotin-labeled probe, but can be adjusted as required.b.Before adding the labeled probe, the reaction mix should be mixed well and incubated at room temperature (20-25℃) for 10 minutes to eliminate possible non-specific binding between probes and proteins...
To above binding reaction, add 1 μl Rgg protein (or buffer as control). To each reaction, add 1 μl fluorescently-labeled DNA probe. Incubate reactions at room temperature (25 °C) for 30 min. While the reactions are incubating, pre-run gel (no samples) at 100 V, 10 min, 4 °...
1. 用于EMSA探针的标记,可以参考如下反应体系:待标记探针(1.75μM)2μlT4 Polynucleotide Kinase 10X Buffer1μlNuclease-Free Water5μl[γ-32P]ATP(3,000Ci/mmol at 10mCi/ml)1μlT4 Polynucleotide Kinase (5-10u/μl)1μl总体积10μl2.如果用于生物素标记,具体方法和步骤可以参考阿拉丁的EMSA探针...
EMSA results using the EBNA control system.Biotin-labeled 60 bp duplex bearing the EBNA-1 binding sequence was incubated with an extract in which the EBNA-1 protein was overexpressed. The binding buffer was supplemented with 50 ng/µl poly(dI•dC), 10% glycerol and 0.05% NP-40. Exposur...
The T4 phage in 1 lac binding buffer (10 mM Tris-HCl, 150 mM KCl, single-stranded binding protein (gp32), was a generous 0.1 mM DTT, and 0.1 mM EDTA). Ten L of diluted DNA with gift from Dr. Peter von Hipel’s laboratory (University of 2 L of 6 EMSA gel-loading solution (...
7、素EMSA: not modified Preparation 2:anneal oligonucleotides 10 x annealing buffer:200 mM Tris 8.0, 10 mM EDTA 8.0, 500 mM NaCl dissolve single strand probe to a concentration of 25uM 50 reaction system:25uM single strand probe 20ul, 10 xannealing buffer:5ul,H2O 5ul 。 put mineral oil...
EMSA+说明书英文版 Non-Radioactive EMSA User Manual VER.3.11 This user manual is suitable for the production SIDET001, SIDET003, SIDET004.Revised 3/2007