Covalently linked complementary oligodeoxynucleotides (splinkers) as tools for molecular biology

MOU*MC«« Self-complementary primer l inkers ("splinkers") speci f ica l ly jo in to restr icted DMA. These oligonucleotides consist of an inverted repeat sequence which forms a double stranded hairpin structure containing a known rest r ic t ion s i te (RS). This bu i l t i n res t r ic t ion s i te enables their specific release from the restr icted DNA fragment. The schematic diagram shows a number of uses for splinker-primed DNA fragments. They can be used as strand specific sequence primers for dideoxy sequencing of either cloned or genomic DNA ( 1 ; e .g . , AMV reverse transcriptase determined sequence of $X174 (T)). This approach also lends i t s e l f to simultaneous strand sequencing by adding unique splinkers to opposite ends. Their specif ic release yields both sequences. Splinker primed fragments can also be completely f i l l e d in and then u t i l i zed as strand specif ic probes ( © ) (coding vs. coding complement) for both Southern and Northern analyses (2) in order to detect unique t ranscr ipts. Furthermore, the addit ion of splinkers to one strand of a DNA fragment automatically imparts a mobi l i ty s h i f t , permitting their use in strand separat ion ( (3 ) ) . Recently, hairpin DNA fragments have been used as l inkers and adapters ( (4 ) ; 3). Thus splinkers have a multitude o.f uses as molecular biological tools. Acknowledgements