O'Neil Casey S, Beach Jacie L, Gruber Todd D
Department of Molecular Biology and Chemistry, Christopher Newport University, Newport News, VA, USA.
Electrophoresis. 2018 Jun;39(12):1474-1477. doi: 10.1002/elps.201700489. Epub 2018 May 4.
DNA gel electrophoresis is a standard tool of biochemistry and molecular biology laboratories. The common dye ethidium bromide suffers from toxicity concerns and requires the use of damaging ultraviolet light. We observe that exposing plasmid DNA to a UV transilluminator for only 1 s results in detectable loss of colonies following transformation, suggesting rapid accumulation of DNA damage. SYBR Safe, a commercial product, is marketed as a safe alternative to ethidium bromide and has excellent sensitivity with nondamaging blue light, but suffers from prohibitively high costs. We show that thiazole orange, the parent compound of SYBR Safe, is an excellent, simple, and inexpensive alternative to these dyes. It is excitable with safe blue light or UV light, with DNA detection limits in agarose gels similar to ethidium bromide and SYBR Safe (1-2 ng/lane). Thiazole orange safely allows the use of nondamaging blue light at the same cost as ethidium bromide.
DNA凝胶电泳是生物化学和分子生物学实验室的标准工具。常用染料溴化乙锭存在毒性问题,且需要使用具有破坏性的紫外线。我们观察到,将质粒DNA暴露在紫外线透射仪下仅1秒,转化后即可检测到菌落损失,这表明DNA损伤迅速积累。商用产品SYBR Safe作为溴化乙锭的安全替代品进行销售,对无损伤蓝光具有出色的灵敏度,但成本高得令人望而却步。我们表明,SYBR Safe的母体化合物噻唑橙是这些染料的一种出色、简单且廉价的替代品。它可用安全蓝光或紫外线激发,在琼脂糖凝胶中的DNA检测限与溴化乙锭和SYBR Safe相似(1-2 ng/泳道)。噻唑橙以与溴化乙锭相同的成本安全地允许使用无损伤蓝光。