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How should ethidium bromide powder be stored after being prepared into a stock solution, and will it affect RNA electrophoresis of porcine adipose tissue?

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Asked by:KitMax 2026-03-20 Views 230 4 answers
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科
科研老司机 2026-09-10
猪脂肪组织总RNA如果上样量低,EB灵敏度会显得勉强,尤其甲醛变性胶背景比普通DNA胶更高。我一般先确保RNA完整性,跑胶前测浓度和A260/A280,上样500-1000 ng总RNA,甲醛变性胶1.2%左右,电泳后用EB后染15-20 min再脱色,通常能看到28S/18S,但低丰度/降解样品可能很弱。如果样品量很少,建议换SYBR Gold或GelRed等灵敏度更高的染料,或先做毛细管电泳QC,不要只靠EB胶判断RNA质量。
E
EmmaAntibody 2026-09-03
反复冻融不推荐。EB母液是水溶液,冻融可能导致局部浓度不均,也可能增加背景或降低有效染色强度。最稳妥的是分装成小体积后4℃避光保存,常用的一管放4℃,其余可以短期存放;不要反复从大瓶里吸液。若发现母液颜色明显变深、出现沉淀或染色背景升高,就不要再用了。对猪脂肪组织RNA这种样品量低、条带比较珍贵的实验,染料状态不好很容易误判,别用跑胶去验证染料。
曾
曾小妹9026 2026-07-15
货号和价格我不好直接报,每个代理商差距大,而且EB粉末/溶液涉及危化品,一周内到货要看本地有没有现货。推荐你直接问Sigma的E8751这类10 mg/mL溶液或粉末规格,让销售确认CAS和运输条件;同时问Thermo的对应10 mg/mL规格,比较交期。猪脂肪组织RNA实验如果这周就要用,别只等进口EB,可以找本地试剂商先拿GelRed或SYBR Safe应急。自己配母液可以省一点,但粉末操作和称量风险更高,实验室不具备条件不建议硬上。
S
SamAntibody 2026-09-18
EB粉末配10 mg/mL母液,一般配好后用棕色瓶或铝箔包住,4℃避光保存。室温短期放一两天问题不大,但长期不建议。配的时候要在通风橱或安全称量区操作,戴口罩、手套,粉末称量要避免扬尘。溶剂可以用灭菌的纯水或PBS,按说明书浓度来,搅拌溶解要充分。配好后建议分装成1 mL小管,避免整瓶反复取用。动物房或公共实验室最好在固定台面操作,配完及时清理台面。
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