Methods and kits for preparing nucleic acid samples
    2.
    发明申请
    Methods and kits for preparing nucleic acid samples 审中-公开
    用于制备核酸样品的方法和试剂盒

    公开(公告)号:US20070020654A1

    公开(公告)日:2007-01-25

    申请号:US11419459

    申请日:2006-05-19

    IPC分类号: C12Q1/68 C12P19/34

    摘要: The present invention provides methods for preparing nucleic acid samples. The methods of the present invention are particularly amenable for preparing samples that substantially represent the whole transcripts. In some aspects the methods include a step of reducing the amount of ribosomal RNA in a total RNA sample prior to amplification. In preferred aspects single stranded sense strand cDNA is generated, labeled and hybridized to arrays of probes. The method is particularly suitable to use with microarray based expression analysis.

    摘要翻译: 本发明提供了制备核酸样品的方法。 本发明的方法特别适于制备基本上代表整个转录物的样品。 在一些方面,所述方法包括在扩增前减少总RNA样品中的核糖体RNA的量的步骤。 在优选的方面,产生单链有义链cDNA,标记并与探针阵列杂交。 该方法特别适用于基于微阵列的表达分析。

    Methods of small sample amplification
    3.
    发明申请
    Methods of small sample amplification 审中-公开
    小样本扩增方法

    公开(公告)号:US20050003392A1

    公开(公告)日:2005-01-06

    申请号:US10805559

    申请日:2004-03-19

    IPC分类号: C12P19/34 C12Q1/68

    CPC分类号: C12P19/34 C12Q1/68

    摘要: The present invention relates to the amplification of nucleic acids, preferably from mRNA. A primer and promoter are added to a target sequence to be amplified and then the target is amplified in an in vitro transcription reaction and the product of this reaction is used as template for subsequent rounds of amplification. Polyadenylated control transcripts are added to the nucleic acid sample prior to the first step of amplification to monitor the efficiency of the amplification and labeling reactions.

    摘要翻译: 本发明涉及核酸,优选mRNA的扩增。 将引物和启动子加入待扩增的靶序列中,然后在体外转录反应中扩增靶,并将该反应的产物用作随后的扩增循环。 将多腺苷酸化的对照转录物在扩增的第一步骤之前加入到核酸样品中,以监测扩增和标记反应的效率。

    Methods for fragmenting nucleic acid
    5.
    发明申请
    Methods for fragmenting nucleic acid 审中-公开
    破碎核酸的方法

    公开(公告)号:US20060141498A1

    公开(公告)日:2006-06-29

    申请号:US11273964

    申请日:2005-11-14

    IPC分类号: C12Q1/68 C12P19/34 C12N9/22

    摘要: Methods for using an apurinic/apyrimidinic endonuclease, capable of cleaving both single- and double-stranded cDNA, for fragmentation and labeling of single stranded or double stranded DNA molecules are provided. Amplification methods that generate single-stranded amplified cDNA are also disclosed. In the subject methods AP sites in a population of nucleic acids are cleaved by an AP endonuclease that is active on both double and single stranded DNA. Fragments may be end labeled. In preferred embodiments APE 1 is used. The methods may be used in a variety of applications where end-labeling single or double stranded DNA is desired.

    摘要翻译: 提供了使用能够切割单链和双链cDNA,用于单链或双链DNA分子的片段化和标记的无嘌呤/脱嘧啶核苷酸内切酶的方法。 还公开了产生单链扩增cDNA的扩增方法。 在本发明方法中,核酸群体中的AP位点被双链和单链DNA上活性的AP内切核酸酶切割。 片段可能是末端标记的。 在优选实施方案中,使用APE 1。 所述方法可用于需要终止标记单链或双链DNA的多种应用。

    Methods for fragmentation and analysis of nucleic acid
    6.
    发明申请
    Methods for fragmentation and analysis of nucleic acid 审中-公开
    核酸分离和分析方法

    公开(公告)号:US20070218478A1

    公开(公告)日:2007-09-20

    申请号:US11612454

    申请日:2006-12-18

    IPC分类号: C12Q1/68 C12P19/34

    摘要: Methods for fragmenting and labeling DNA in a single reaction volume and incubation step using a uracil DNA glycosylase, an apurinic/apyrimidinic endonuclease, and a terminal transferase are disclosed. In a preferred embodiment the UDG, AP and TdT activities are first mixed together to form an enzyme mixture and then the enzyme mixture is mixed with the uracil containing DNA. The fragmentation and labeling reactions thus take place simultaneously as part of the same reaction. The methods may be used in a variety of applications where fragmenting and end-labeling single or double stranded DNA is desired.

    摘要翻译: 公开了在单个反应体积中分离和标记DNA的方法以及使用尿嘧啶DNA糖基化酶,无嘌呤/脱嘧啶核苷酸内切酶和末端转移酶的温育步骤。 在优选的实施方案中,首先将UDG,AP和TdT活性混合在一起以形成酶混合物,然后将该酶混合物与含有尿嘧啶的DNA混合。 因此,碎裂和标记反应同时作为相同反应的一部分进行。 该方法可用于需要片段化和末端标记单链或双链DNA的多种应用。

    Method for depleting specific nucleic acids from a mixture
    7.
    发明申请
    Method for depleting specific nucleic acids from a mixture 审中-公开
    从混合物中消耗特定核酸的方法

    公开(公告)号:US20050123987A1

    公开(公告)日:2005-06-09

    申请号:US11040759

    申请日:2005-01-21

    IPC分类号: C12P19/34 C12Q1/68

    摘要: The presently claimed invention provides methods, compositions, and apparatus for analyzing nucleic acids isolated from blood. Specifically, the present invention provides a method of analyzing blood samples by blocking amplification of selected unwanted RNAs and subsequently analyzing the amplified sample by hybridization to a plurality of probes attached to a solid support. In one embodiment, the invention provides enriching for a population of interest in a complex population by diminishing the presence of an unwanted sequence that may interfere with the analysis of sequences of interest.

    摘要翻译: 本发明提供了用于分析从血液中分离的核酸的方法,组合物和装置。 具体地,本发明提供了通过阻断所选择的不想要的RNA的扩增来分析血液样品的方法,并且随后通过与连接到固体支持物的多个探针杂交来分析扩增的样品。 在一个实施方案中,本发明通过减少可能干扰感兴趣序列分析的不想要的序列的存在来提供对复杂群体中的感兴趣群体的富集。