ISSN: 0973-7510

E-ISSN: 2581-690X

Parveen Kumar , Pronab Dhar, Vikramaditya Upmanyu, Amit Kumar and Ashok Kumar Tiwari
1Division of Biological Standardization, Indian Veterinary, Research Institute, Izatnagar -243122, India.
J Pure Appl Microbiol. 2015;9(2):1185-1188
© The Author(s). 2015
Received: 06/02/2015 | Accepted: 10/03/2015 | Published: 30/06/2015
Abstract

In this study, a 6.5 Kb fragment of cell culture adapted Lapinized Classical Swine Fever Virus (CSFV) was cloned into pTZ57R/T vector using the TA molecular cloning technique. Briefly, viral RNA was isolated using a modified RNA isolation protocol using Ribozol and RNeasy followed by cDNA synthesis using Clontech superscript RT. The cDNA obtained was used to synthesize 6.5 kb PCR products using (Long acting) LA-PCR. Gel purified PCR product was cloned with pTZ57R/T vector which was confirmed by RE digestion and partial sequencing. The same protocol can be utilized for cloning of similar sized amplified products in molecular biology laboratories.

Keywords

Classical swine fever, LA-PCR, TA Cloning

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