BACKGROUND Accurate quantification of mRNA in whole blood is made difficult by the simultaneous degradation of gene transcripts and unintended gene induction caused by sample handling or uncontrolled activation of coagulation. This study was designed to compare a new blood collection tube (PAXgene Blood RNA System) and a companion sample preparation reagent set with a traditional sample collect...

when duplicate blots were probed with the avidin-peroxidase conjugate (Figure 1B, lanes 5′–8′) ExtrAvidin® (Sigma, St. Louis, MO, USA). Unlike the secondary antibody conjugate used in Figure 1A, the avidin-peroxidase conjugate used in Figure 1B did not recognize the electrophoresed immunoglobulins within the immunoprecipitate. Moreover, specific bands of ezrin were readily detected in immunopre...

Cytochrome P450 2C19 (CYP2C19) is the enzyme responsible for the metabolism of a number of drugs, including anticonvulsants and antidepressants. We have developed a semi-quantitative competitive RT-PCR assay to estimate the degree of expression of the full-length CYP2C19 message. This assay used a known quantity of internally deleted CYP2C19 RNA to quantitate the RT-PCR products of the CYP2C19 ...

Isolation of high-quality RNA free of contaminants, such as polyphenols, proteins, plant secondary metabolites, and genomic DNA from plant tissues, is usually a challenging but crucial step for molecular analysis. We developed a novel protocol based on the cetyltrimethylammonium bromide method to isolate high-quality RNA from blackberry plant tissues, especially fruits. Most DNA was remove...

Isolation of pure and intact RNA is essential, but the concentration and integrity of total RNA preparations is affected by the purification process used. Applications for which RNA will ultimately be used may include reverse transcriptase (RT)-PCR, preparations of targets for microarrays, ribonuclease protection assays, preparations of cDNA libraries and northern blotting. Recently, the import...

in this study, several designed creb1-specific shrnas cloned into lentiviral transfer vectors were screened in293t by transient transfection assays. relative creb1 expression was determined by relative qrt-pcr. all of the designed shrnas gave some level of knockdown. one shrna was more effective for knocking down the specific gene among others. the predicted sirnas activities as determined by r...

A 4-day-old neonate born prematurely at 29 weeks gestation developed thrombocytopenia (platelet count 43 10/L) with associated severe pulmonary and intracranial hemorrhaging. Urgent transfusions with 10 mL/kg of packed red cells and 10 mL/kg of platelet concentrate were given. These were sourced from blood donated 2 days earlier in Singapore by a 21-yearold female university student who was cli...