All the single-stranded oligonucleotides of the Td listed in Table S1 were synthesized by Sangon Biotech Co. (Shanghai, China). The Luria-Bertani culture (LB, pH = 7.4) was bought from Sinopharm (Beijing, China), HEPES (pH = 7.4), NaCl, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) was bought from Sigma-Aldrich LLC. (Shanghai, China). Biotin, quinine sulfate, N-hydroxysulfosuccinimide (NHS) were provided by Tianjin Macklin (Shanghai, China). streptavidin-functionalized magnetic beads (SA−MBs) with a diameter of 2–3 μm were purchased from BaseLine ChromTech Research Centre (Tianjin, China). DNA loading buffer (6×) and DNA Markers were provided by Sangon Biotechnology Co., Ltd. (Shanghai, China). All solutions with a resistivity of 18.2 MΩ cm were prepared using a Milli-Q water purification system (Sartorius, Germany).
Dna loading buffer 6
Manufactured by Sangon
Sourced in China
DNA loading buffer (6×) is a laboratory reagent used to prepare DNA samples for gel electrophoresis. It is a concentrated solution that increases the density of the DNA sample, allowing it to sink into the gel. The buffer also contains tracking dyes to visually monitor the progress of the electrophoresis.
Automatically generated - may contain errors
3 protocols using dna loading buffer 6
1
Sensitive DNA Oligonucleotide Detection Protocol
2
Polyacrylamide Gel Electrophoresis of CRISPR Complexes
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For the polyacrylamide gel experiment, lanes 1–4 from left to right were loaded with 500 nM G1-B-10 strand and 500 nM G1-SE+-T-19 strand, 750 nM LbaCas12a and 500 nM gRNA (lane 1); 500 nM G1-B-11 strand and 500 nM G1-SE+-T-20 strand (lane 2); 500 nM G1-B-11 strand and 500 nM G1-SE+-T-19 strand (lane 3); and 500 nM G1-B-10 strand and 500 nM G1-SE+-T-19 strand (lane 4). To a 200 μl PCR tube, the above mix, 7.5 μl NEBuffer2.1 (10×) and 4 mM Mg2+ were added and brought up to a total volume of 50 μl by deionized water. The reaction process was 1 h, followed by 95°C for 30 min, 55°C for 5 min and then 37°C for 10 min. Then, DNA loading buffer (6×) (Sangon Co., China) was added to the tube. A 15% denatured polyacrylamide gel was used to separate the DNA strands. Electrophoresis was carried out at 100 V for 2 h. The gel was then removed and soaked in the dye solution (4S Red Plus Nucleic Acid Stain, Sangon Co.) for 30 min. The gel image was taken by Bio-rad ChemDoc XRS (Bio-Rad, USA).
3
Oligonucleotide Synthesis and Purification
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The oligonucleotides
listed inTable S1 were synthesized and
purified by Sangon Biotech Co., Ltd. (Shanghai, China). Native-PAGE
gel and DNA loading buffer (6×) were purchased from Sangon Biotech
Co., Ltd. (Shanghai, China). Ultrapure water was obtained from a Millipore
water purification system.
listed in
purified by Sangon Biotech Co., Ltd. (Shanghai, China). Native-PAGE
gel and DNA loading buffer (6×) were purchased from Sangon Biotech
Co., Ltd. (Shanghai, China). Ultrapure water was obtained from a Millipore
water purification system.
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