小中大不知道这种方法与上两种方法比较怎么样？兄弟可以把这个产品的实验方法发下或者把该公司的说明书影印一份，供大家参考。或者您以上方法都用过，希望给个参考信息。
我的本意是提供一个集中的关于MiRNA的平台，大家可以在这里互通有无。所以希望广大有经验的战友多多提供意见以及帮助，而不是光有我在这里班门弄斧。
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这种方法是一个叫做MiraMas的kit中所使用的，制造商是美国的Bioo Scientific公司。我给它起的名字叫通用接头连接法：
在RT步骤，该kit通过试剂盒内含的特殊ligase连接5’端腺苷化且3’端封闭的寡核苷酸接头(Adenylated 3’ Ligation Adapter)于所有miRNA的3’末端，然后通过与该接头序列互补的3’端引物经M-MLV反转录酶将所有miRNA同步反转录成cDNA。
主要优点：
1.比加A尾法更高的反转录效率：
"The advantage of adding the 3' adapter sequence using RNA ligase as used in MiraMas kit, provides higher specificity for priming the reverse transcription step and also the binding site for the Reverse PCR primer only at the 3' ends of the RNAs, compared to using polyA polymerase to add homopolymeric A's followed by hybridization of the oligo dT/adapter to provide the binding sites for RT and Rev PCR primer. The problem with the polyA approach is that the oligo dT can hybridize to internal homopolymeric A regions in mRNAs, instead of only to 3' end."
2.更多的单样品miRNA检测数量：
"Each MiraMas reaction can be used to make cDNA library to allow for analysis of 10's - 100's of microRNA targets (depending on mass amount of input RNA), compared to Qiagen kit each reaction only makes enough cDNA for around 5 target microRNAs. And Qiagen kit does not allow flexibility to use high or low input amounts of RNA. Using the low input amounts, researcher can process lots of samples from one kit."
