中國海洋大學(xué)基因工程l4第二章pcr技術(shù)-文庫吧資料
2024-09-24 20:32本頁面
【正文】 containing a suitable overhang at one end. PCR amplification of the uncharacterized sequence X is then possible using a primer specific for the known DNA sequence (A) and a primer specific for one of the strands of the bubble in the vectorette linker (B). The vectorette linker primer B cannot prime DNA synthesis initially as there is no sequence to which it can bind: it is identical, not plementary in sequence to one strand of the bubble, and unrelated in sequence to the other. However, primer A initiates synthesis of a plementary DNA strand which will contain a sequence plementary to one of the unique sequences within the bubble linker. As a result, primer B can bind to this newly synthesized strand and initiate new strand synthesis to start a PCR reaction. The flanking sequence Y can similarly be isolated in another reaction using a suitable Aspecific primer and a bubblespecific primer derived from the strand opposite to that used for making primer B. 3 Figure . AluPCR permits amplification of DNA sequences located between two closely positioned but oppositely orientated Alu repeats. (A) Primers can be designed from the Alu consensus sequence to hybridize to sequences at either terminus of the repeat element, with the 5 direction pointing away from the repeat element. (B) A single such primer can be used in a PCR reaction to amplify sequences located between two Alu repeats which are in opposite orientation, and arranged such that the two bound primers are convergent (. PCR with primer 2 alone can amplify sequence X but not Y or Z。雙聚合酶 消除錯(cuò)配 ） or合成能力強(qiáng)酶 (Tth) ? *Tricine（三羥甲基甘胺酸） pH穩(wěn)定性 ? PCR buffer ? 、 KCl ? dNTP
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