发明名称 |
Characterising cDNA comprising cutting sample cDNAs with a first endonuclease, sorting fragments according to the un-paired ends of the DNA, cutting with a second endonuclease then sorting the fragments |
摘要 |
A method for characterising cDNA, which comprises: (a) cutting a sample comprising a population of one or more cDNAs or isolated fragments thereof, each having a strand complementary to the 3' poly-A terminus of an mRNA and bearing a tail, with a first sampling endonuclease at a first sampling site of known displacement from a reference site proximal to the tail to generate from each cDNA or isolated fragment thereof a first and second sub-fragment, each comprising a sticky end sequence of predetermined length and unknown sequence, the first sub-fragment bearing the tail; (b) sorting either the first or second sub-fragments into sub-populations according to their sticky end sequence and recording the sticky end sequence of each sub-population as the first sticky end; (c) cutting the sub-fragments in each sub-population with a second sampling endonuclease, which is the same as or different from the first sampling endonuclease, at a second sampling site of known displacement from the first sampling site to generate from each sub-fragment a further sub-fragment comprising a second sticky end sequence of predetermined length and unknown sequence; and (d) determining each second sticky end sequence; wherein the aggregate length of the first and second sticky end sequences of each sub-fragment is from 6 to 10; and wherein the sequences and relative positions of the reference site and first and second sticky ends characterise the or each cDNA.
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申请公布号 |
NZ334426(A) |
申请公布日期 |
2000.09.29 |
申请号 |
NZ19970334426 |
申请日期 |
1997.09.05 |
申请人 |
BRAX GROUP LIMITED |
发明人 |
SCHMIDT, GUNTER;THOMPSON, ANDREW HUGIN |
分类号 |
C12N15/09;C12N15/10;C12Q1/68;(IPC1-7):C12Q1/68 |
主分类号 |
C12N15/09 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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