background: erythropoietin (epo) is a glycoprotein hormone function to regulate the production of red blood cells. deficiency of epo is known to cause anemia in chronically infected renal patients and they require regular blood transfusion. availability of recombinant epo has eliminated the need for blood transfusion and now it is extensively used for the treatment of anemia. glycosylation of e...

Background: Hepatitis delta virus (HDV) is a subviral human pathogen that exploits host RNA editing activity to produce two essential forms of the sole viral protein, hepatitis delta antigen (HDAg). Editing at the amber/W site of HDV antigenomic RNA leads to the production of the large form (L-HDAg), which is required for RNA packaging. Methods: In this study, PCR-based site-directed mutagen...

Overlap syndrome, which is known as the coexistence of chronic obstructive pulmonary disease (COPD) and obstructive sleep apnea (OSA), was first defined by Flenley. Although it can refer to concomitant occurrence of any of the pulmonary diseases and OSA, overlap syndrome is commonly considered as the coexistence of OSA and COPD. This disease has unique adverse health consequences distinct from ...

BACKGROUND Radioactive DNA polymerase activity methods are cumbersome and do not provide initial extension rates. A simple extension rate assay would enable study of basic assumptions about PCR and define the limits of rapid PCR. METHODS A continuous assay that monitors DNA polymerase extension using noncovalent DNA dyes on common real-time PCR instruments was developed. Extension rates were ...

background human telomerase reverse transcriptase (htert) has become an ideal target for development of anticancer therapy. small interfering rnas (sirnas) are very powerful reagents for gene silencing and show promise for cancer gene therapy. however, only a small number of sirnas have been demonstrated to be effective. for gene therapy targeting htert, it is essential to develop a robust syst...

The ability to edit the genome is essential for many state-of-the-art experimental paradigms. Since DNA breaks stimulate repair, they can be exploited to target site-specific integration. The clustered, regularly interspaced, short palindromic repeats (CRISPR)/cas9 system from Streptococcus pyogenes has been harnessed into an efficient and programmable nuclease for eukaryotic cells. We thus com...

Molecular cloning methods based on primer and overlap extension PCR are widely used due to their simplicity, reliability,low cost and high efficiency. In this article, an efficient mega primer (MP)-mediated cloning strategy for chimeragenesis and long DNA fragment insertion is presented. MP cloning is a seamless, restriction/ligation-independent method that requires only three steps: i) the fir...