Isolation of High Quality RNA from Cereal Seeds Containing High Levels of Starch.

12:23 EDT 29th May 2015 | BioPortfolio

Summary of "Isolation of High Quality RNA from Cereal Seeds Containing High Levels of Starch."


INTRODUCTION:
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.
OBJECTIVE:
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.
METHODOLOGY:
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.
RESULTS:
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.
CONCLUSION:
The study has described an easy, efficient and highly reproducible method for RNA isolation from various cereal seeds.

Affiliation

Shanghai Key Laboratory of Bio-Energy Crops, School of Life Sciences, Shanghai University, No. 333 Nanchen Road, Shanghai, P.R. China.

Journal Details

This article was published in the following journal.

Name: Phytochemical analysis : PCA
ISSN: 1099-1565
Pages:

Links

PubMed Articles [36254 Associated PubMed Articles listed on BioPortfolio]

Comparison of RNA Isolation Methods From Insect Larvae.

Isolating RNA from insects is becoming increasingly important in molecular entomology. Four methods including three commercial kits RNeasy Mini Kit (Qiagen), SV Total RNA isolation system (Promega), T...

Gene expression of antioxidant enzymes and coffee seed quality during pre- and post-physiological maturity.

Seeds collected at different maturation stages vary in physiological quality and patterns of protective antioxidant systems against deterioration. In this study we investigated the expression of genes...

Characterization of mustard seeds and paste by DART ionization with time-of-flight mass spectrometry.

Direct analysis in real time (DART) is a novel technique with great potential for rapid screening analysis. The DART ionization method coupled with high-resolution time-of-flight mass spectrometry (TO...

Selection of process parameters for producing high quality defatted sesame flour at pilot scale.

The present work was undertaken to study the effect of pearling duration, soaking time, steaming duration and drying temperature on the quality of sesame seeds and mechanically extracted partially def...

Measuring Risk-Adjusted Value Using Medicare and ACS-NSQIP: Is High-Quality, Low-Cost Surgical Care Achievable Everywhere?

To evaluate the relationship between risk-adjusted cost and quality for colectomy procedures and to identify characteristics of "high value" hospitals (high quality, low cost).

Clinical Trials [6642 Associated Clinical Trials listed on BioPortfolio]

Fibre and Appetite Regulation Trial (FART)

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 Prostate Brachytherapy as Salvage for Locally Recurrent Prostate Cancer Previously Treated With External Beam Radiotherapy

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...

Low Glycemic Index Diets (With Pulses) in Type 2 Diabetes

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...

High-Dose Brachytherapy in Treating Patients With Prostate Cancer

This phase I/II trial studies the side effects and how well high-dose brachytherapy works in treating patients with prostate cancer that has not spread to other parts of the body. Brachyth...

Protein, Fiber, and Metabolic Syndrome - The PROFIMET Study

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 ...

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.

Search BioPortfolio:
Loading
Advertisement

Relevant Topic

Food
Latest News Clinical Trials Research Drugs Reports Corporate
Food is any substance consumed to provide nutritional support for the body. It is usually of plant or animal origin, and contains essential nutrients, such as carbohydrates, fats, proteins, vitamins, or minerals. The substance is ingested by an organism ...

Advertisement

Searches Linking to this Article