Enzyme-free cloning: a rapid method to clone PCR products independent of vector restriction enzyme sites

Enzyme-free cloning: a rapid method to clone PCR products independent of vector restriction enzyme sites.

We describe a simple method for the cloning of PCR products without the need for post-amplification enzymatic treatment. Tailed PCR primer sets are used to create complementary staggered overhangs on both insert and vector by a post-PCR denaturation-hybridisation reaction. The single-stranded overhangs are designed to allow directional cloning in a ligase-free manner. This 'enzyme-free cloning'...

Rapid Restriction Enzyme-Free Cloning of PCR Products: A High-Throughput Method Applicable for Library Construction

Herein, we describe a novel cloning strategy for PCR-amplified DNA which employs the type IIs restriction endonuclease BsaI to create a linearized vector with four base-long 5'-overhangs, and T4 DNA polymerase treatment of the insert in presence of a single dNTP to create vector-compatible four base-long overhangs. Notably, the insert preparation does not require any restriction enzyme treatmen...

High-efficiency T-vector cloning of PCR products by forced A tagging and post-ligation restriction enzyme digestion.

simple and rapid identification of most medically important candida species by a pcr-restriction enzyme method

opportunistic fungal infections including candidiasis have increased dramatically in recent years. most medically important fungi are candida species. rapid identification od candida isolates to the species level in the clinical laboratory is necessary for more rapid and effective antifungal therapy and to facilitate hospital infection control measures. conventional morphological methods for id...

Simple and Rapid Identification of Most Medically Important Candida Species by a Pcr- Restriction Enzyme Method

Opportunistic fungal infections including candidiasis have increased dramatically in recent years. Most medically important fungi are Candida species. Rapid identification of Candida isolates to the species level in the clinical laboratory is necessary for more rapid and effective antifungal therapy and to facilitate hospital infection control measures. Conventional morphological methods for id...