نتایج جستجو برای: cloning efficiency
تعداد نتایج: 450287 فیلتر نتایج به سال:
background: the homologous recombination (hr) is one of the conventional cloning methods for the production of recombinant dna. it is a quick method for in vivo dna cloning without using the high cost restriction enzymes. a few modifications in the cloning procedure can increase the efficiency of this method. m aterials and methods: in this study, effect of heating on the rate of the igg1 heavy...
Somatic cell nuclear transfer (SCNT) is an assisted reproductive technique that can produce multiple copies of excellent livestock. However, low cloning efficiency limits the application of SCNT. In this study, we systematically investigated the major influencing factors related to the overall cloning efficiency in pigs. Here, 13620 cloned embryos derived from excellent pigs were transferred in...
Background Despite successful cloning of many mammalian species, the cloning efficiency is extremely low compared to that of IVF raising the possibility of the existence of epigenetic barrier preventing successful cloning. MaterialsAndMethods Using comparative transcriptome analysis comparing transcriptomes of IVF and SCNT embryos and that of donor cells, we identified epigenetic barrier and fi...
human diploid fibroblast cells produce a spectrum of necessary growth factors and extracellular matrix (ecm) components essential for growth and proliferation of a variety of other cell types. in this study, the effect of five human embryonic fibroblast cell lines, isolated from liver, lung, skin and foreskin tissues, was investigated. a coculture system analyse was employed to cloning efficien...
Simple and low-cost recombinant enzyme-free seamless DNA cloning methods have recently become available. In vivo Escherichia coli cloning (iVEC) can directly transform a mixture of insert and vector DNA fragments into E. coli, which are ligated by endogenous homologous recombination activity in the cells. Seamless ligation cloning extract (SLiCE) cloning uses the endogenous recombination activi...
The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3' adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3' overhanging thymidine. The aim of this work was to offer in a single vector the ability to clone both types of PCR products. For that p...
We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP furth...
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