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Recombinant DNA technology
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
A cDNA library is a population of bacterial transformants or phage lysates in which each mRNA isolated from an organism or tissue is represented as its cDNA insertion in a plasmid or a phage vector. The frequency of a specific cDNA in such a library would ordinarily depend on the frequency of the concerned mRNA in the tissue/organism in question. This enzyme performs similar reactions as DNA polymerase and has an absolute requirement for a primer with a free 3′-OH. When eukaryotic mRNA is used as a template, a poly-T oligonucleotide (more specifically, oligodeoxynucleotide) is conveniently used as the primer since these mRNAs have a poly-A tail at their 3′-ends. However, special tricks are required to utilize primers for other RNAs such as prokaryotic mRNA, rRNA, and RNA virus genomes.
Recombinant DNA Technology
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2020
cDNA Library: A cDNA library is a combination of cloned cDNA (complementary DNA) fragments inserted into a collection of host cells, which constitute some portion of the transcriptome of the organism and are stored as a library (Figure 4.13).
The green-synthesized curcumin-mediated zinc oxide nanoparticles (CmZnO-NP) as the exclusive antioxidant and efficient wound healing agent compared with curcumin, methanol, phenytoin, and ZnO
Published in Inorganic and Nano-Metal Chemistry, 2021
Elahe Dianati, Vida Hojati, Jina Khayatzadeh, Saeedeh Zafar Balanezhad
The skin tissue slabs of the wound area in different treated groups of rats were homogenized. The homogenization process was conducted in presence of 2ME and PBS buffer to prevent RNA content degradation. The tissue RNA content was extracted utilizing an RNA extraction kit (Norgen Biotek Corp. Canada). The tissue cDNA library was prepared by reverse transcribing the tissue total RNA content using a reverse transcription kit (Qiagen, Hilden, Germany). Then, the primer sets of wound healing gene markers including Collagen type II and VEGF as the target genes were designed by Allel-ID6 software (Table 2). THE real-time PCR technique was utilized by applying an SYBR green-supplemented PCR master mix (Ampliqon, Denmark). Finally, the PCR efficiency and GAPDH expression (hose-keeping gene) were measured and normalized, respectively.