The ratio of absorbance at 260 and 280 nm (the A260/280 ratio) is frequently used to assess the purity RNA DNA preparations. Data presented in this report demonstrate significant variability when different sources water were perform spectrophotometric determinations. Adjusting pH for analysis from approximately 5.4 a slightly alkaline 7.5-8.5 significantly increased ratios 1.5 2.0. Our studies revealed that changes both ionic strength solution influenced ratios. In addition, ability detect...

In this report, we present DNAzol, a patent-pending DNA isolation reagent containing guanidine thiocyanate and detergent mixture. It is complete, nontoxic ready-to-use for the of genomic from various biological sources. DNAzol protocol, sample homogenized (or lysed) in precipitated with ethanol, washed dissolved 8 mM NaOH. Following pH adjustment, can be used immediately analysis or stored at 4 degrees C. The entire completed 20-30 min, wide range molecules isolated including fragments down...

Journal Article Solubilzation in formamide protects RNA from degradation Get access Piotr Chomczynski University of Cincinnai College Medicine231 Bethesda Avenue, Cincinnati, OH 45267–0547, USA Search for other works by this author on: Oxford Academic PubMed Google Scholar Nucleic Acids Research, Volume 20, Issue 14, 25 July 1992, Pages 3791–3792, https://doi.org/10.1093/nar/20.14.3791 Published: 1992 history Received: 06 May

The role of the pituitary, and in particular, GH GH-releasing hormone (GRH) gene expression was studied hypophysectomized rats with without treatment. Hypothalamic GRH mRNA 6-fold greater than control rats. Increased levels were observed at 3 days maximal increase noted 7 days, postoperatively. Administration to hypox partially reversed mRNA, suggesting a negative feedback regulation by activity transcript accumulation level. overall expression, however, appears more complex since content...

We have isolated and cloned the full length cDNA for mouse GH-releasing hormone (mGRH) from hypothalamus using a recently described strategy involving polymerase chain reaction technique (PCR). Degenerate oligonucleotide primers were selected based on short (six amino acids) conserved regions in human rat GRH peptides that would recognize DNA sequences encoding similar acids regardless of codon usage. Primer-extended was amplified by PCR templates prepared reverse transcribing total...

Growth hormone (GH) has been implicated in the pathogenesis of proliferative diabetic retinopathy. We sought to determine whether this could be mediated by an effect GH on proliferation endothelial cells, and, for purpose, established long-term cultures human retinal microvascular cells (hREC) from normal postmortem eyes. High-purity (greater than 95%) hREC preparations were selected experiments, based immunofluorescence with acetylated low density lipoprotein (LDL) and anti-factor...

Insulin is essential for the accumulation of rat casein messenger RNA (mRNA) in presence glucocorticoid and prolactin. The certain mRNA's other tissues has also been linked to insulin action. present study shows that effect on 25,000 molecular weight mRNA does not reflect stabilization transcript by insulin. Rather, its synthesis

Relatively little is known about the range of RNA levels in human blood. This report provides assessment peripheral blood level and its inter-individual differences a group 35 healthy humans consisting 25 females 10 males ranging age from 50 to 89 years. In this group, average total was 14.59 μg/ml blood, with no statistically significant difference between males. The individual ranged 6.7 22.7 subjects, repeated sampling an individual's showed that level, whether high or low, stable. can be...

Three different methods for RNA preparation using guanidine are presented in this unit--a single-step isolation method employing liquid-phase separation to selectively extract total from tissues and cultured cells two that rely on a CsCl step gradient isolate RNA.

The role of thyroid hormone and GH in the regulation hypothalamic GH-releasing (GRH) gene expression rat was examined after induction deficiency by thyroidectomy. Thyroidectomy resulted a time-dependent decrease GRH content, which significant 2 weeks postoperatively, reduction pituitary content to 1% control level 4 weeks. In contrast, secretion incubated hypothalami under both basal K+-stimulated conditions increased Hypothalamic mRNA levels also exhibited timedependent increase, at 1 week...