The Prime It RmT random primer labeling kit uses random oligonucleotides as primers for labeling DNA to produce high-specific-activity probes.The procedure relies on the ability of random 9-mers to anneal to multiple sites along the length of a DNA template. The primer–template complexes formed represent a substrate for the magenta DNA polymerase, a thermostable polymerase that allows the Stratagene Prime-It RmT random primer labeling kit to be stored at room temperature. The enzyme synthesizes new DNA by incorporating nucleotide monophosphates at the free 3´-OH group provided by the primer. The newly synthesized DNA is made radioactive by substituting radiolabeled [α-32P]dCTP for unlabeled dCTP in the reaction mixture. The resulting labeled DNA serves as a sensitive hybridization probe for screening gene libraries,4 probing Southern and Northern blots,5,6 and in situ hybridization techniques.
Qualified reagents for the polyadenylation of microRNAs (miRNAs) from total RNA and synthesis of first-strand cDNA from the tailed miRNAs. Detects mature miRNA. Differentiate miRNA that differ by a single nucleotide. Sensitive detection down to 10 copies. Highly accurate results in 3 hours miRNA. Specific Forward Primers miRNA Primer Design Guidelines
Random oligonucleotides have been used as primers in a method for labeling DNA to produce high specific-activity probes.1,2 The procedure relies on the ability of random hexanucleotides to anneal to multiple sites along the length of a DNA template. The primer–template complexes formed represent a substrate for the Klenow fragment of DNA polymerase I. The enzyme synthesizes new DNA by incorporating nucleotide monophosphates at the free 3´–OH group provided by the primer. The newly synthesized DNA is made radioactive by substituting a radiolabeled nucleotide for a nonradioactive one in the reaction mixture. The resulting labeled DNA can serve as a sensitive hybridization probe for screening gene libraries,3 probing Southern and northern blots,4,5 and in situ hybridization techniques.
Qualified reagents for the polyadenylation of microRNAs (miRNAs) from total RNA and synthesis of first-strand cDNA from the tailed miRNAs. Detects mature miRNA Differentiate miRNA that differ by a single nucleotide Sensitive detection down to 10 copies Highly accurate results in 3 hours miRNA Specific Forward Primers miRNA Primer Design Guidelines For Research Use Only. Not for use in diagnostic procedures.
