Orange ruler 10 bp dna ladder

To determine the length of the DNA molecules, an agarose gel electrophoresis was performed. A 3% agarose gel was produced by dissolving 1.2 g of agarose in 40 mL of 1x TAE-Buffer (50x Tris/Acetic Acid/EDTA (TAE), diluted with distilled deionized water) by heating in a microwave oven. For staining of the DNA, 4 μL nucleic acid gel stain were added to the gel. After casting the gel, it was allowed to solidify for 30 min and was put into a Mini-Sub cell GT cell from BIO RAD. For the preparation of the DNA samples, 5 μL of a DNA loading dye were added to 25 μL of sample. Twelve microliters of the samples and 5 μL of Orange Ruler 10 bp DNA ladder (Thermo Fischer, USA) were pipetted on the gel and allowed to run for about 30 min at 120 V. Gels were afterwards imaged using a ChemiDocTM MP imaging system from BIO RAD. For the measurement of the DNA content, an Implen NanoPhotometer® was used. Therefore, the photometer was blanked, using only MES-buffer and afterwards the samples were measured in triplicates at 260 nm.

Gel electrophoresis was
used to quantify JD-DNA conjugation yield as well as aptamer hybridization
efficiency. Gels were prepared with 4% SeaKem LE agarose (Lonza) dissolved
in 1× Tris-Borate-EDTA (TBE) buffer. SYBR Gold Nucleic Acid Gel
Stain (1×) (Thermo Fisher Scientific) was added to the gel prior
to polymerization. JD-DNA samples containing 2 μg of DNA in
0.1 M TEAA were prepared with 6× Orange Gel Loading Dye (New
England Biolabs, Inc.) and loaded into the gel. The O’RangeRuler
10 bp DNA ladder (Thermo Fisher Scientific) was used to approximate
DNA size. Gels were electrophoresed at 180 V and then imaged using
the NuGenius gel documentation workstation (Syngene). Densitometry
analysis was performed using ImageJ software (National Institutes
of Health).