公开(公告)号：EP1552018B1

公开(公告)日：2009-07-01

申请号：EP03784781.1

申请日：2003-07-18

申请人： NEW ENGLAND BIOLABS, INC.

发明人： TZERTZINIS, George ,  FEEHERY, George ,  TUCKEY, Corinna ,  NOREN, Christopher ,  MCREYNOLDS, Larry

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12N15/111 ,  C12N2310/14 ,  C12N2330/30 ,  C12N2330/31 ,  C12Y301/26003

公开(公告)号：EP2470675B1

公开(公告)日：2016-03-30

申请号：EP10812579.0

申请日：2010-08-25

申请人： New England Biolabs, Inc.

发明人： BITINAITE, Jurate ,  VAISVILA, Romualdas ,  PRADHAN, Sriharsa ,  ZHENG, Yu ,  ROBERTS, Richard, J. ,  CHIN, Hang-gyeong ,  COHEN-KARNI, Devora ,  NOREN, Christopher ,  RALEIGH, Elisabeth, A. ,  WILSON, Geoffrey G.

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6883 ,  C12Q1/6827 ,  C12Q2600/154 ,  C12Q2521/331 ,  C12Q2525/191 ,  C12Q2531/113

公开(公告)号：EP2470675A1

公开(公告)日：2012-07-04

申请号：EP10812579.0

申请日：2010-08-25

申请人： New England Biolabs, Inc.

发明人： BITINAITE, Jurate ,  VAISVILA, Romualdas ,  PRADHAN, Sriharsa ,  ZHENG, Yu ,  ROBERTS, Richard, J. ,  CHIN, Hang-gyeong ,  COHEN-KARNI, Devora ,  NOREN, Christopher ,  RALEIGH, Elisabeth, A. ,  WILSON, Geoffrey G.

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6883 ,  C12Q1/6827 ,  C12Q2600/154 ,  C12Q2521/331 ,  C12Q2525/191 ,  C12Q2531/113

摘要： Methods and compositions are described for detecting hydroxymethylated nucleotides (hmNs) in a polynucleotide preparation with a view to mapping the location of hmNs in a genome, quantifying the occurrence of hmNs at selected loci and correlating the occurrence of hmNs with gene expression and phenotypic traits. Embodiments describe the use of modifying enzymes together with site-specific endonucleases to detect the hmNs.

摘要翻译： 描述了用于检测多核苷酸制备物中的羟甲基化核苷酸（hmNs）的方法和组合物，以便映射hmNs在基因组中的位置，定量选择的基因座处hmNs的发生并将hmNs的发生与基因表达和表型性状相关联。 实施方案描述了将酶与位点特异性核酸内切酶一起修饰以检测hmNs的用途。

公开(公告)号：EP1552018A2

公开(公告)日：2005-07-13

申请号：EP03784781.1

申请日：2003-07-18

申请人： NEW ENGLAND BIOLABS, INC.

发明人： TZERTZINIS, George ,  FEEHERY, George ,  TUCKEY, Corinna ,  NOREN, Christopher ,  MCREYNOLDS, Larry

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12N15/111 ,  C12N2310/14 ,  C12N2330/30 ,  C12N2330/31 ,  C12Y301/26003

摘要： A method for obtaining a mixture of heterogeneous short double-stranded RNA molecules suitable for use in gene silencing (hsiRNA) by subjecting large double-stranded RNA to enzymatic cleavage under specified conditions. The resulting mixture consistently includes enhanced representation of fragments having a size of 21-22 nucleotides absent any fractionation step. The fragments contain sequences that collectively span the entire length of the large double-stranded RNA from which they are derived. Double-stranded RNA with sequences that individually represent segments of a target mRNA may be analyzed using the methods described herein to identify the most active subset of hsiRNA fragments or individual siRNA fragments for achieving gene silencing for any gene or transcribed sequences. A method is additionally provided for preparing and cloning DNA encoding selected siRNA, hsiRNA mixtures or hairpin sequences to provide a continuous supply of a gene silencing reagent derived from any long double-stranded RNA.