Oligo dT primer and method for constructing cDNA library

A DNA molecule and library construction technology, applied in the field of cDNA library construction and Oligo dT primers, can solve the problems of single library construction method, large operation restrictions, and small scope of application

Active Publication Date: 2015-03-04
盛司潼
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Problems solved by technology

[0013] The object of the present invention is to provide a kind of Oligo dT primer, solve the Oligo dT primer in the prior art that the scope of application is small, based on the Oligo dT primer in the prior art, the operation restriction after forming double-stranded cDNA molecules is large, and the library construction method is applied single question

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  • Oligo dT primer and method for constructing cDNA library
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  • Oligo dT primer and method for constructing cDNA library

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[0084] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0085] The type IIs restriction endonuclease of the present invention is a restriction endonuclease whose cutting site is outside the recognition sequence, including but not limited to: AcuI, AlwI, BbsI, BbVI, BccI, BceAI, BciVI, BfuAI, BmrI , BpmⅠ, BpuEⅠ, BsaⅠ, BseMII, BseRI, BsgⅠ, BsmAⅠ, BsmBI, BsmFI, BspCNⅠ, BspMI, BspQⅠ, BtgZⅠ, EarⅠ, EciⅠ, EcoP15Ⅰ, FauⅠ, FokⅠ, HgaⅠ, HphⅠ, HpyAV, MboⅡ, MlyⅠ, MmeⅠ, MnlⅠ , NmeAIII, PleI, SapI, SfaNI and TspDTI, preferably AcuI, BsgI, EcoP15I or MmeI.

[0086] The mRNA of the present invention is derived from eukaryotic organisms, including but not limited to animals, plants, fungi and protists.

[0087] An Oligo dT primer, the Oligo dT primer is a single-stranded DNA molecule containing a continuous thymidine deo...

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Abstract

The invention relates to the field of molecular biology and provides an Oligo dT primer. The Oligo dT primer contains single-chain DNA molecules having a continuous dT sequence, wherein the DNA molecules contain recognition sites used for cutting; and the recognition sites used for cutting refer to uridine monophosphate, deoxyinosine or phosphorothioate bond. The invention also provides a method for constructing a cDNA library on the basis of the Oligo dT primer. The Oligo dT primer disclosed by the invention is high in applicability and can be applied to multiple different library construction schemes; the initiation sites at the polyA tails of mRNA molecules can be accurately positioned; and according to the method for constructing the cDNA library disclosed by the invention, all the mRNA molecules in samples can be specifically reflected in the constructed cDNA library.

Description

[0001] This case was filed on February 28, 2012, the application number is 201210047909.3, and the title of the invention is a divisional application of "A Oligo dT Primer and a Method for Constructing a cDNA Library". technical field [0002] The invention relates to the field of genetic engineering, and more specifically relates to an Oligo dT primer and a method for constructing a cDNA library based on the Oligo dT primer. Background technique [0003] The analysis of gene expression level plays a crucial role in the study of gene function. Gene expression sequence analysis (serial analysis of gene expression, SAGE) is a technology for rapid analysis of gene expression information, which finds out expression SAGE tag sequences with different abundances, so as to obtain genome expression information nearly completely. SAGE technology and gene chip are currently the two most common gene expression profiling methods. With the development of second-generation sequencing te...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C40B40/08C40B50/06
Inventor 盛司潼
Owner 盛司潼
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