The recent development of real-time PCR has offered the opportunity of sensitive and accurate quantification of mRNA levels that is crucial in biomedical research. Although reverse transcription (RT)-PCR is at present the most sensitive method available, many low abundant mRNAs are, although detectable, often not quantifiable. Here we report an improved two-step real-time RT-PCR procedure using...

Real-time PCR is an accurate method that can be used for the quantification of specific DNA molecules. Here we provide a protocol for SYBR Green I in real-time PCR applications using plastic reaction tubes. We report that SYBR Green I is alkali labile and once degraded inhibits the PCR. In our optimized protocol, diluted aliquots of SYBR Green I remain stable for at least two weeks. We also eva...

BACKGROUND Global analysis of the genome, transcriptome, and proteome is facilitated by the recent development of tools for large-scale, highly parallel analysis. We describe a novel nucleic acid amplification system that generates products by several methods. 3'-Ribo-SPIA primes cDNA synthesis at the 3' polyA tail, and whole transcript (WT)-Ribo-SPIA primes cDNA synthesis across the full lengt...

A cDNA library was constructed from firefly (Photinus pyralis) lantern poly(A)+ RNA, using the Escherichia coli expression vector lambda gt11. The library was screened with anti-P. pyralis luciferase (Photinus luciferin:oxygen 4-oxidoreductase, EC 1.13.12.7) antibody, and several cDNA clones expressing luciferase antigens were isolated. One clone, lambda Luc1, contained 1.5 kilobase pairs of cD...

background: recent researches have demonstrated that human t-cell immunoglobulin mucin 1 (tim-1) glycoprotein plays important roles in regulation of autoimmune and allergic diseases, as well as in tumor immunity and response to viral infections. therefore,targeting tim-1 could be a potential therapeutic approach against such diseases.objectives: in this study, we aimed to express tim-1 protein ...

We describe an approach to crowdsource the evaluation of TTS systems by preference tests and report on lessons learnt from running 127 real-life crowdsourced tests. We show that at least one type of cheating becomes more prevalent over time if left unchecked and develop metrics to exclude cheaters. We demonstrate that their exclusion improves test outcomes.

Cervical cancer is the one of leading cause death for women worldwide. It occurs due to infection with human papilloma virus (HPV) which attacks cervix in female reproductive organs. The detecting cervical method by using pap smears and liquid based cytology (LBC) vaccinations. However, both methods are limited quickly accurately. This study aims find potential biomarkers that will be used dete...

Amplification of the 5' ends of cDNA, although simple in theory, can often be difficult to achieve. We describe a novel method for the specific amplification of cDNA ends. An oligo-dT adapter incorporating a dUTP-containing PCR primer primes first-strand cDNA synthesis incorporating dUTP. Using the Cap finder approach, another distinct dUTP containing adapter is added to the 3' end of the newly...