Dna ladder

Gel electrophoresis was conducted to confirm the on-chip amplification as follows. A 2% agarose (Bioneer) solution was prepared by microwaving with 30 s pulses for 3 min. A 2 μL aliquot of 10 mg/mL ethidium bromide (EtBr) (Sigma-Aldrich) was added to 50 mL of a 2% agarose solution to visualize the DNA under UV light, and the mixture was solidified in a template. The prepared gel was placed into the gel chamber and filled with 1× TBE (Bioneer) containing 5 μL of 10 mg/mL EtBr. A 2 μL aliquot of eluted RNA from the chip was mixed with 6 μL of loading buffer (Sigma-Aldrich) and loaded carefully along with the DNA ladder (Sigma-Aldrich) into the lanes of the gel. Gel electrophoresis was conducted at 80 V for 1 h until the dye line was approximately 80% of the way down the gel. The gel was carefully placed inside a UV box (Gel documentation system, GDS-200D) to take a photographic image.

Maxime RT Premix Kit was purchased from INtRon Biotechnology (Catalog # 25082, Korea). DNA Ladder (Catalog # P1473), DEPC-treated water (clear colorless liquid) (Catalog #D5758), Triton^™X100 (liquid colorless to light yellow, CAS #9002-93-1), STZ (white to yellow powder, CAS #18883-66-4) and the primer sequences (dried in 2 ml screw cap tube) were obtained from Sigma-Aldrich, USA. Antibodies were purchased from Novus Biologicals, USA. MT as Glucophage tablets (500 mg white colored, film-coated tablets) and GI as Amaryl (2 mg light pink colored oval shaped, uncoated tablets) were purchased from Eva Pharma, Egypt. Solvents and other associated biochemical reagents were obtained with high grade from Sigma-Aldrich, USA.

Magnesium chloride hexahydrate (MgCl₂⋅6H₂O), sodium chloride (NaCl), acetone, and phosphate buffer saline (PBS) were purchased from Sinopharm Chemical Reagent Co., Ltd. (China). Exonuclease III was provided by Thermo Fisher Scientific Co., Ltd. (United States). Tris borate EDTA (TBE) buffer (5×), tris EDTA (TE) buffer (1×), sodium dodecyl sulfate (SDS, 10%, w/v) buffer, loading dye buffer solutions (6×), agarose, and saline-sodium citrate (SSC, 20×) buffer solutions were obtained from Solarbio Science & Technology Co., Ltd. (China). GelRed Neuclic Acid Gel Stain was purchased from Biotium, Inc. (United States). DNA ladder was provided by Sigma-Aldrich Co., Ltd. (China). QIAamp DNA Blood Mini Kit was purchased from Qiagen Co., Ltd. (Germany). All chemicals in our experiments were of analytical grade and used without further purification. Aqueous solutions were prepared using deionized water (≥18 MΩ, Milli-Q, Millipore). The SERS substrates were provided by Nanova Biomaterials Inc. (United States). High-performance liquid chromatography (HPLC)-purified oligonucleotides were provided by Sangon Biotechnology Co., Ltd. (China).