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    • 4. 发明授权
    • Method of sequencing DNA
    • DNA测序方法
    • US06210891B1
    • 2001-04-03
    • US09269436
    • 1999-07-06
    • Pål NyrenMathias UhlenMostafa Ronaghi
    • Pål NyrenMathias UhlenMostafa Ronaghi
    • C12Q168
    • C12Q1/6869C12Q2565/518C12Q2565/301C12Q2535/101
    • The present invention provides a method of identifying a base at a target position in a single-stranded sample DNA sequence wherein an extension primer, which hybridizes to the sample DNA immediately adjacent to the target position, is provided and the sample DNA and extension primer are subjected to a polymerization reaction in the presence of a deoxynucleotide or dideoxynucleotide, whereby the deoxynucleotide or dideoxynucleotide will only become incorporated and release pyrophosphate if it is complementary to the base in the target position. Release of pyrophosphate is detected enzymatically and pyrophosphate detection enzyme(s) are included in the polymerization step.
    • 本发明提供了在单链样品DNA序列中鉴定目标位置的碱基的方法,其中提供了与紧邻目标位置的样品DNA杂交的延伸引物,并且样品DNA和延伸引物为 在脱氧核苷酸或双脱氧核苷酸的存在下进行聚合反应,由此脱氧核苷酸或双脱氧核苷酸仅在靶位置与碱基互补时才被引入并释放焦磷酸盐。 检测到焦磷酸盐的释放,聚合步骤中包括焦磷酸盐检测酶。
    • 10. 发明授权
    • Chemical method for the analysis of DNA sequences
    • 用于分析DNA序列的化学方法
    • US5534424A
    • 1996-07-09
    • US331552
    • 1995-04-18
    • Mathias UhlenJoakim Lundeberg
    • Mathias UhlenJoakim Lundeberg
    • C12N15/09C12Q1/68C12P19/34C07H21/04C12Q1/70
    • C12Q1/6858C12Q1/6834C12Q1/6869
    • The invention provides a method of identification of the base in a target position in a DNA sequence wherein sample DNA is subjected to amplification; the amplified DNA is immobilised and then subjected to strand separation, the non-immobilised strand being removed and an extension primer, which hybridises to the immobilised DNA immediately adjacent to the target position, is provided; each of four aliquots of the immobilised single stranded DNA is then subjected to a polymerase reaction in the presence of a dideoxynucleotide, each aliquot using a different dideoxynucleotide whereby only the dideoxynucleotide whereby only the dideoxynucleotide complementary to the base in the target position becomes incorpored; the four aliquots are then subjected to extension in the presence of all four deoxynucleotides, whereby in each aliquot the DNA which has not reacted with the dideoxynucleotide is extended to form double stranded DNA while the dideoxy-blocked DNA remains as non-extended DNA; followed by identification of the double stranded and/or non-extended DNA to indicate which dideoxynucleotide was incorporated and hence which base was present in the target position.
    • PCT No.PCT / EP93 / 01203 371日期1995年04月18日 102(e)日期1995年4月18日PCT提交1993年5月12日PCT公布。 公开号WO93 / 23562 日期:1993年11月25日。本发明提供了在DNA序列中的靶位置鉴定碱基的方法,其中对样品DNA进行扩增; 将扩增的DNA固定化,然后进行链分离,除去非固定化链和延伸引物,其与紧邻目标位置的固定化DNA杂交; 然后在双脱氧核苷酸存在的情况下,将四个等分的固定化单链DNA中的每一个进行聚合酶反应,每个等分试样使用不同的双脱氧核苷酸,从而只有双脱氧核苷酸,其中只有与目标位置的碱基互补的双脱氧核苷酸才会被吸收; 然后在四种脱氧核苷酸的存在下将四个等分试样延伸,由此在每个等分试样中,未与双脱氧核苷酸反应的DNA延伸形成双链DNA,而双脱氧封闭的DNA保留为非延伸的DNA; 随后鉴定双链和/或未延伸的DNA以指示哪个双脱氧核苷酸被引入,因此哪个碱基存在于目标位置。