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Leiothrix lutea karyogene primers and sequencing method and application thereof

A nuclear gene and sequencing technology, which is applied in the field of Acacia red-billed nuclear gene primers and their sequencing, can solve problems such as non-amplification and non-specific amplification.

Active Publication Date: 2017-05-24
ANHUI NORMAL UNIV
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Problems solved by technology

[0005] In the prior art, the use of universal primers to amplify the nuclear gene sequence may result in failure to amplify or non-specific amplification

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  • Leiothrix lutea karyogene primers and sequencing method and application thereof
  • Leiothrix lutea karyogene primers and sequencing method and application thereof

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Embodiment Construction

[0028] In the following, the present invention will be described in further detail through the description of the embodiments, so as to help those skilled in the art have a more complete, accurate and in-depth understanding of the inventive concepts and technical solutions of the present invention.

[0029] The sequencing method of the red-billed acacia nuclear gene primer of the present invention comprises the following steps:

[0030] (1) Carry out the extraction of total genome to red-billed lovesickness bird, extraction method adopts phenol, chloroform extraction method, concrete steps are as follows:

[0031] 1) Cut about 100mg of muscle tissue sample, shred it and place it in a sterilized 2.0ml EP tube;

[0032] 2) Add 1ml of DNA extraction solution and 4ul of 20mg / ml RNaseA (ribonuclease) into the EP tube, shake well and place in a water bath at 37°C for 1 hour;

[0033] 3) Take out the EP tube, add 5ul 20mg / ml proteinase K, shake well, put it in a water bath at 50°C f...

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Abstract

The invention discloses leiothrix lutea karyogene primers and a sequencing method and an application thereof. The primers comprise a first primer pair, a second primer pair and a third primer pair; the first primer pair comprises RAG-F1-17 having a sequence number of SEQ ID NO.1 and RAG-R1-533 having a sequence number of SEQ ID NO.2; the second primer pair comprises RAG-F2-996 having a sequence number of SEQ ID NO.3 and RAG-R2-2024 having a sequence number of SEQ ID NO.4; the third primer pair comprises RAG-F3-1945 having a sequence number of SEQ ID NO.5 and RAG-R3-2779 having a sequence number of SEQ ID NO.6. The obtained primers have quite good amplification function and are used for amplification of a leiothrix lutea karyogene rag1, finally, bright strips can be obtained, and follow-up study of large-scale use of the primers for amplification is facilitated.

Description

technical field [0001] The invention belongs to the technical field of bird phylogeny, and in particular relates to a red-billed acacia nuclear gene primer and a sequencing method and application thereof. Background technique [0002] The coding products of nuclear recombination-activating genes (RAGs), rag1 and rag2, constitute the most important recombinases, which can specifically recognize the RSS on both sides of the V, DH, and J gene fragments of Ig and TCR, and catalyze DNA double-strand breaks, causing V (D) J rearrangement, the main cause of diversity in Ig and TCR repertoires. The missense mutation of rag1 or rag2 can partially lose the activity of the encoded recombinase, and the tendency of V(D)J recombination will change, resulting in the blockage of early lymphocyte development and the loss of T and B cells in the circulation, resulting in A type of severe combined immunodeficiency, clinically known as Omenn syndrome (OS). At present, genes are mainly used as...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2531/113C12Q2565/125
Inventor 阚显照王莹陈仁瑞章勤蒋澜王青青吴璇丁恒武易双龙宛凤英王会梅
Owner ANHUI NORMAL UNIV
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