生物样本首先在含有异硫氰酸胍的变性缓冲液中裂解并匀浆,DNase和RNase立即失活,可确保DNA和RNA的完整性。将裂解物通过AllPrep DNA Mini纯化柱。在高盐缓冲液中,纯化柱与基因组DNA选择性高效结合。在优化的缓冲液条件下进行柱上Proteinase K消化,可从各种类型样本中纯化获得高产量DNA。之后洗脱获得即用型DNA。在AllPrep DNA Mini纯化柱的流出液中加入乙醇,用Proteinase K进行消化。消化后再次加入乙醇,包括miRNA在内的总RNA结合到RNeasy Mini纯化柱上。用DNase I消化获得高产量RNA,且不含DNA。洗涤步骤可冲走污染物,之后可洗脱获得高品质RNA。
AllPrep操作流程仅需75分钟,即可快速、便利的纯化获得高品质核酸,可即用于多种下游应用(参见"The fast and convenient AllPrep procedure")。该试剂盒使用创新的试剂,因此无需使用苯酚等有毒物质,极大减少了对环境和使用者的危害。AllPrep DNA/RNA/miRNA Universal Kit可在QIAcube全自动核酸纯化仪上实现自动化,确保流程标准化,为您带来更大的便利。
To visualize contamination of RNA with genomic DNA, quantitative real-time PCR assays were performed with (+RT) or without (–RT) reverse transcriptase. Unlike kits from other suppliers, the AllPrep DNA/RNA/miRNA Universal Kit delivered high yields of pure RNA from difficult-to-lyse adipose tissue, as demonstrated by c-Jun expression assays.