TOOLS UltraFast PCR Cloning Kit

TOOLS UltraFast PCR Cloning Kit

TOOLS UltraFast PCR cloning kit is designed for fast and directional cloning of the single or multiple DNA into any vector at any site, regardless of DNA fragment end compatibility. This kit is applicable for efficient clone of fragments of 50 bp - 10 kb. The vector is linearized at the cloning site. A small sequence overlapped with each end of the cloning site is added onto the insert through PCR. The insert and the linearized vector, with overlapped sequences of 15 bp - 20 bp on both 5’- and 3’-end, respectively, are mixed in an appropriate ratio and incubated with recombinase 2X UltraFast Mix at 50°C for 5 - 15 min. This kit is a novel cloning kit, applicable for homologous recombination with 1 - 5 fragments. The Kit is independent of DNA ligase, significantly reducing the self-ligated colonies. The enhanced recombinase and highly optimized buffer included in 2X UltraFast Mix significantly improve the recombination efficiency and the tolerance to impurities. pFAST vector is compatible with most PCR products, enabling the specific PCR products to be used directly for recombination without any treatments, which significantly simplify the procedure.

產品型號 規格 單價 庫存 購買數量
TTC-CA15 25 rxns. -
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  1. Fast: Recombination cloning only 5 minutes

  2. Multiple cloning:Homologous recombination with 1-5 fragments

  3. High efficiency: insert size 50bp~10kb, >95% positive clone

  4. Flexible: Directly clone into your expression vector and replace TA cloning ( pFAST vector included)

實驗數據

UltraFast Cloning Kit Competition Result:

常見問題

Q1.Ultra Fast Cloning kit是否可以同時將2kb、8kb、2kb三段insert放進Vector當中?

A:由於其Insert長度>10kb,需再討論一下,針對insert序列先看一下,可以嘗試看看,其中8kb的部分需使用效果好的Polymerase進行放大

 

Q2.TOOLS UltraFast PCR Cloning Kit操作的時候,Vector和Insert是否都需要再做Clean up的動作?

A:不用,但依照Protocol說明,需要先將限制酶酵素失活,以免影響後續的實驗。

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