Dulbecco’s phosphate-buffered saline (dPBS, 2.6 mMKCl, 1.47 mM KH2 PO4, 137 mMNaCl, and 8.05 mM Na2HPO4), Hanks’ Balanced Salt Solution (HBSS--,no calcium, no magnesium, refered as HBSS) were obtained from Thermo Fisher Scientific (Waltham, MA). Invitrogen Novex™ TBE-Urea Gels, 15%, 10 well, and TBE-Urea Sample Buffer (2X), for denaturing conditions, and Novex™ TBE Running Buffer (5X), and Novex TBE Gels, 4–20% (non-denaturing conditions) were obtained from Thermo Fisher Scientific. Gel Loading Dye, purple (6X), without SDS was obtained from New England Biolabs (Ipswich, Massachusetts). Five hundred nanometer streptavidin beads were purchased from Bangs Laboratories (Fishers, IN). MiRCURY LNA miRNA Inhibitors (antimiRs) were obtained from Qiagen (Germantown, MD).
Novex tbe running buffer
Manufactured by Thermo Fisher Scientific
Novex™ TBE Running Buffer is a pre-made, concentrated solution used for electrophoresis of nucleic acids in TBE buffer systems. It is designed to be diluted with water prior to use.
Automatically generated - may contain errors
Lab products found in correlation
Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions)
Novex tbe gel, by Thermo Fisher Scientific (1 mentions)
Mircury lna mirna inhibitor, by Qiagen (1 mentions)
Dulbecco s phosphate buffered saline, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Sybr gold nucleic acid stain, by Thermo Fisher Scientific (1 mentions)
Novex hi density tbe sample buffer, by Thermo Fisher Scientific (1 mentions)
Geldoc go imaging system, by Bio-Rad (1 mentions)
Fla 5100 fluorescent image analyser, by Fujifilm (1 mentions)
3 protocols using novex tbe running buffer
1
Denaturing Gel Electrophoresis of miRNA
2
miRNA mimic Stability in FBS
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20 μl of 5 μM miRNA mimic was mixed with 80 μl of FBS (Gibco #10270) and incubated at 37°C. Samples were taken at 0 and 24 h time points. 20 pmol of each sample was loaded in Novex™ Hi-Density TBE Sample buffer (Thermofisher) and analyzed on Novex™ 20% TBE Gel. The gel was run for 50 min at 250 V in Novex™ TBE Running Buffer (Thermofisher). After electrophoresis, gels were incubated in Sybr™ Gold Nucleic Acid Stain (Thermofisher) for 10 min and subsequently washed twice with water. Imaging was done with UV transillumination using the GelDocGo imaging system (Biorad).
3
Oligonucleotide Binding Assay Protocol
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Self-annealed oligonucleotide C (at a concentration of 200 nM) was pre-incubated with the indicated dilution series of each complex, in BUFFER D for a period of 30 min at 4°C. NativePAGE 4× loading buffer (Thermo Fisher Scientific) was then added to each sample, before being applied to 0.8% (w/v) tris–borate–EDTA (TBE)–agarose gel. Electrophoresis was carried out by application of 50 V for a period of 3 h at 4°C, in 0.5% (v/v) TBE (Novex TBE Running Buffer, Thermo Fisher Scientific). Separated species were visualised using a Fuji FLA-5100 Fluorescent Image Analyser, using excitation with a 473 nm laser.
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