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    • 5. 发明申请
    • SELECTIVE TAGGING OF SHORT NUCLEIC ACID FRAGMENTS AND SELECTIVE PROTECTION OF TARGET SEQUENCES FROM DEGRADATION
    • 选择性标记短核酸片段和选择性保护目标序列从降解
    • WO2010115044A3
    • 2011-03-31
    • PCT/US2010029690
    • 2010-04-01
    • FLUIDIGM CORPZIMMERMANN BERNHARD G
    • ZIMMERMANN BERNHARD G
    • C12Q1/68C12N15/10C12N15/11
    • C12N15/1093C12N15/10C12N15/1065C12N15/1072C12Q1/6846C12Q2521/307C12Q2521/325C12Q2537/159
    • Methods are provided for selective tagging of short nucleic acids comprising a short target nucleotide sequence over longer nucleic acids comprising the same target nucleotide sequence. The methods can involve performing one or two cycles of amplification of a sample comprising long nucleic acids and short nucleic acids, each comprising the same target nucleotide sequence with at least two target-specific primers or primer pairs under suitable annealing conditions, wherein the primer pairs comprise: an inner primer or primer pair that can amplify the target nucleotide sequence on long and short nucleic acids (wherein each inner primer comprises a 5' nucleotide tag; and an outer primer or primer pair that amplifies the target nucleotide sequence on long nucleic acids, but not on short nucleic acids); whereby the amplification after a second cycle produces at least one tagged target nucleotide sequence that comprises two nucleotide tags, one from each inner primer, with the target nucleotide sequence located between the nucleotide tags.
    • 提供了用于在包含相同靶核苷酸序列的较长核酸上选择性标记包含短目标核苷酸序列的短核酸的方法。 所述方法可以包括在合适的退火条件下进行包含长核酸和短核酸的样品的一个或两个循环的扩增,每个循环包含相同的靶核苷酸序列与至少两个靶特异性引物或引物对,其中引物对 包括:可以扩增长和短核酸上的靶核苷酸序列的内部引物或引物对(其中每个内引物包含5'核苷酸标签;以及扩增长核酸上的靶核苷酸序列的外引物或引物对 ,但不在短核酸上); 由此在第二周期之后的扩增产生至少一个标记的靶核苷酸序列,其包含两个核苷酸标签,每个内引物中的一个,靶核苷酸序列位于核苷酸标签之间。