Isolation of High Quality RNA from Cereal Seeds Containing High Levels of Starch.
Summary of "Isolation of High Quality RNA from Cereal Seeds Containing High Levels of Starch."
Cereals are an important source of food, feed and fuel with a rapidly increasing global demand. However, cereal seeds contain high levels of starch and polysaccharides, making the isolation of high quality RNA extremely difficult.
To develop a novel method for extracting high quality total RNA from various starch- and polysaccharides-rich cereal seeds, such as maize, rice, sorghum and wheat.
We developed a modified sodium dodecyl sulphate (SDS)/TRIzol method. The combined use of a Tris buffer (pH 9.0) and SDS before TRIzol extraction effectively resolved the problem of seed homogenate solidification in such a buffer. A high concentration of SDS was used separately, not only to promote cell lysis but also to effectively dissolve seed sample containing high levels of starch. Moreover, acid phenol saturated with 0.1 m citrate buffer (pH 4.3) was used to separate RNA from DNAs, proteins and high levels of starch. This rapid protocol was compared with other RNA isolation methods preferentially used for plants rich in polysaccharides and secondary metabolites.
Gel electrophoresis analysis indicated that the extracted total RNA had good integrity without apparent DNA contamination. Furthermore, an A(260/280) ratio of approximately 2.0, an A(260/230) ratio of more than 2.0 and RIN values of more than 8.6 indicated that the isolated RNA was of high purity. The isolated RNA was suitable for subsequent molecular manipulations, such as reverse-transcription polymerase chain reaction (PCR), rapid amplification of cDNA ends (RACE) and real-time PCR.
The study has described an easy, efficient and highly reproducible method for RNA isolation from various cereal seeds.
Shanghai Key Laboratory of Bio-Energy Crops, School of Life Sciences, Shanghai University, No. 333 Nanchen Road, Shanghai, P.R. China.
This article was published in the following journal.
Name: Phytochemical analysis : PCA
The study of gene expression in maize varieties represents a powerful tool aiming to increase vitamin A precursors. However, the isolation of RNA from different maize varieties is challenge because th...
Quality improvement of cereal brans, a health promoting ingredient for functional foods is the emerging research concept due to their low shelf stability and presence of non-nutrient components. A stu...
We dramatically improved a plasmid-isolation protocol based on the popular alkaline-sodium dodecyl sulfate plasmid isolation method. Our modified method provides significant time and cost savings. We...
Data quality is one of the major concerns of using crowdsourcing websites such as Amazon Mechanical Turk (MTurk) to recruit participants for online behavioral studies. We compared two methods for ensu...
Coffee (Coffea arabica L.) seeds are sensitive to desiccation and oxidative stress during drying processes. We investigated the effect of drying and moisture levels on germination-related gene express...
High intake of cereal fibre has been shown to be associated with reduced weight gain and improved insulin sensitivity. We hypothesize these effects are due to the short chain fatty acids...
High dose rate (HDR) brachytherapy is a form of radiation treatment using temporary radioactive seeds. This is done by placing very tiny catheters or tubes into the prostate and then inser...
Healthy individuals with type 2 diabetes will receive intensive counseling on food selection to improve glucose control using either high cereal fiber dietary strategies or low glycemic in...
This is a randomized controlled single-blinded intervention study in 111 overweight and obese subjects with risk factors of developing type 2 diabetes, with the aim to investigate effects...
The size of the prostate changes due to insertion of the radioactive seeds. We will measure the prostate size, before and after the procedure.
Medical and Biotech [MESH] Definitions
A plant genus of the family POLYGONACEAE that is used as a cereal grain. Although the seeds are used as cereal, the plant is not one of the cereal grasses (POACEAE).
A method of producing a high-quality scan by digitizing and subtracting the images produced by high- and low-energy x-rays.
Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.
Ventilatory support system using frequencies from 60-900 cycles/min or more. Three types of systems have been distinguished on the basis of rates, volumes, and the system used. They are high frequency positive-pressure ventilation (HFPPV); HIGH-FREQUENCY JET VENTILATION; (HFJV); and high-frequency oscillation (HFO).
Abnormalities in the serum levels of LIPIDS, including overproduction or deficiency. Abnormal serum lipid profiles may include high total CHOLESTEROL, high TRIGLYCERIDES, low HIGH DENSITY LIPOPROTEIN CHOLESTEROL, and elevated LOW DENSITY LIPOPROTEIN CHOLESTEROL.