摘要 |
The invention provides ligation reaction mixtures, methods, and kits for improved template-independent in-tramolecular ligation (circularization) of linear ssDNA, including denatured gDNA fragments or first-strand cDNA made by re-verse transcription of RNA, using, for example, a thermostable RNA ligase. The circular ssDNA molecules obtained using the im-proved ligation reaction mixtures and methods can be used, for example, as templates: for amplification by inverse PCR, rolling circle replication, transcription, or for massively parallel DNA sequencing. Applications include, for example: gene expression analysis by qPCR or using microarrays; analysis of gDNA copy number variation; and detection or quantification of specific nu-cleic acid sequences for research, screening, medical diagnostics, theranostics, personalized medical treatment or breeding, for purposes such as human or animal medicine, forensics, or agriculture. |