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GRiSP Research Solutions - PCR & qPCR, RNA Research, and Sample Purification

GC05.0020 - Xpert TA Easy Cloning Kit, 20 reactions

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The Xpert TA Easy cloning kit is designed for the direct ligation of PCR fragments containing 3´-A overhangs, generated using non-proofreading DNA polymerases such as Taq or blends thereof, into a ready-to-use stable linearized vector. The vector is prepared by digestion of pGRS TA Easy cloning vector with EcoRV and the subsequent addition of a single thymidine at each of the 3´-ends. This system offers greater efficiency than most TA vectors available with over 700 positive colonies under optimal conditions. The system has a low background of less than 4% due to reduced self-ligation.

The pGRS TA Easy vector (3015bp) contains the β-lactamase gene resulting in ampicillin resistance for selection purposes. The PCR amplicon is inserted between pUC/M13 primers, which facilitates DNA sequencing. Flanking Lac and dual opposed SP6/T7 promoters ensure the possibility for correct in vitro transcription of the insert, independently of its orientation. Flanking EcoRI and NotI recognition sites enable single enzyme digestion for subcloning.

Features

  • High efficiency with over 700 positive colonies
  • Low background due to self-ligation
  • Ampicillin resistant

Applications

  • Direct ligation of PCR fragments containing 3´-A overhangs into a ready-to-use stable linearized vector

Contents

  • pGRS TA Easy cloning vector (50ng/µl), 20 reactions
  • T4 DNA Ligase (5 Weiss U/µl), 100 U
  • T4 DNA Ligase Buffer (5X), 200µl
  • Control Insert (600bp), 5µl