| 摘要 |
<p>PURPOSE: A composition for hot-start PCR is provided to prevent DNA band smearing by primer-dimer formation or non-specific reaction. CONSTITUTION: A composition for hot-start PCR contains a blocking oligonucleotide which is complementary to a primer and has blocked hydroxyl group at 3' end. The melting temperature of the blocking oligonucleotide is lower than the primer having the complementary base sequence. The hydroxyl group site of the blocking oligonucleotide is substituted with substituent except for hydroxyl group. The substituent is C3-Space, C6-space, C12-space, C18-Space, amine, phosphate, DIG, or thiol. The composition additionally contains dye substance which is non-reactive to a template DNA. The dye substance is bromophenol blue, xylene cyanol, bromocresol, or cresol red. [Reference numerals] (AA) PCR reactant mixing step, reverse transcription step; (BB) PCR step; (CC, DD) Template; (EE) Primer + blocking primer; (FF) Primer; (GG) Blocking oligo; (HH) No amplification, DNA polymerase inactivation; (II) Amine; (JJ) Polymerase; (KK) Primer + blocking primer, denaturation; (LL) Blocking oligo; (MM) Denaturation; (NN) Annealing; (OO) Blocking oligo separation; (PP) Amplification; (QQ) Elongation</p> |
