Compositions Including A Double Stranded Nucleic Acid Molecule And A Stem-Loop Oligonucleotide

Abstract

The present invention concerns preparation of DNA molecules, such as a library, using a stem-loop oligonucleotide. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.

1 - 54 . (canceled) 
     
     
         55 . A kit comprising:
 a stem-loop oligonucleotide comprising an inverted repeat and a loop, wherein the stem loop oligonucleotide further comprises at least one deoxy-uridine;   a ligase; and   a polymerase.   
     
     
         56 . The kit according to  claim 55 , wherein the kit further comprises deoxy-uridine glycosylase. 
     
     
         57 . The kit according to  claim 55 , wherein the kit further comprises an endonuclease. 
     
     
         58 . The kit according to  claim 55 , wherein the kit further comprises a thermostable polymerase. 
     
     
         59 . The kit according to  claim 55 , wherein the kit further comprises and dNTPs. 
     
     
         60 . The kit according to  claim 55 , wherein the ligase is a DNA ligase. 
     
     
         61 . The kit according to  claim 55 , wherein the polymerase is a DNA polymerase. 
     
     
         62 . The kit according to  claim 55 , wherein the kit further comprises a universal primer. 
     
     
         63 . A kit comprising:
 a stem-loop oligonucleotide comprising an inverted repeat and a loop, wherein the stem loop oligonucleotide further comprises at least one deoxy-uridine;   a DNA ligase; and   a DNA polymerase.   
     
     
         64 . The kit according to  claim 63 , wherein the kit further comprises deoxy-uridine glycosylase. 
     
     
         65 . The kit according to  claim 63 , wherein the kit further comprises an endonuclease. 
     
     
         66 . The kit according to  claim 63 , wherein the kit further comprises a thermostable polymerase. 
     
     
         67 . The kit according to  claim 63 , wherein the kit further comprises and dNTPs. 
     
     
         68 . The kit according to  claim 63 , wherein the kit further comprises a universal primer. 
     
     
         69 . A kit comprising:
 a stem-loop oligonucleotide comprising an inverted repeat and a loop, wherein the stem loop oligonucleotide further comprises at least one deoxy-uridine;   a DNA ligase;   a DNA polymerase;   deoxy-uridine glycosylase; and   an endonuclease.   
     
     
         70 . The kit according to  claim 69 , wherein the kit further comprises a thermostable polymerase. 
     
     
         71 . The kit according to  claim 69 , wherein the kit further comprises and dNTPs. 
     
     
         72 . The kit according to  claim 69 , wherein the kit further comprises a universal primer.