均一化说明书.pdfVIP

P01011 EasyClone normalized cDNA library construction package User Manual Page 1 of 47 Introduction Introduction The EasyClone normalized cDNA library construction package is designed to synthesize full-length-enriched double stranded (ds) cDNA from total or polyA+ RNA, followed by normalization of amplified cDNA and subcloning into an appropriate library vector . By following the instructions provided in this manual, you will be able to construct highly complex normalized cDNA libraries which can be used in various applications, including yeast two-hybrid screening , DUALmembrane or DUALhunter screening or expression screening . Page 2 of 47 Kit contents Kit contents The EasyClone normalized cDNA library construction package provides reagents for 20 cDNA synthesis reactions . The kit comprises following adapter pair: CDS-3M and PlugOligo-3M. This pair is suitable for synthesis of cDNA for directional cloning. BOX 1 Component Amount 5x First-Strand Buffer 80 µl DTT (20 mM) 30 µl 10x dNTP mix (10 mM each) 110 µl PlugOligo-3M adapter (15 µM)* 25 µl 5‘-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGGGG-P-3‘ CDS-3M adapter (10 µM)* 25 µl 5‘-AAGCAGTGGTATCAACGCAGAGTGGCCGAGGCGGCC(T)20VN -3‘ EasyClone Reverse Transcriptase