Advertisement

Topics

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

06:00 EDT 9th July 2011 | 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

DeepDyve research library

PubMed Articles [36096 Associated PubMed Articles listed on BioPortfolio]

Canonical correlations between agronomic traits and seed physiological quality in segregating soybean populations.

The objective of this study was to evaluate the relationship between agronomic traits and physiological traits of seeds in segregating soybean populations by canonical correlation analysis. Seven popu...

An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis.

Plant seeds accumulate large amounts of storage reserves comprising biodegradable organic matter. Humans rely on seed storage reserves for food and as industrial materials. Gene expression profiles ar...

Production of high-quality islets from goettingen minipigs: Choice of organ preservation solution, donor pool, and optimal cold ischemia time.

The transplantation of porcine islets into man might soon become reality for patients with type 1 diabetes mellitus. Therefore, porcine islets of high quality and quantity, and a scalable isolation pr...

A rapid and high-quality method for total RNA isolation from Haematococcus pluvialis.

Haematococcus pluvialis, as the most potential natural source of astaxanthin, which is a powerful antioxidant with high economic value, has attracted more and more scientific attention in recent years...

High quality colonoscopy must be secured Colonoscopy plays a key role in the work-up of digestive diseases and constitutes the cornerstone in colorectal cancer diagnosis and prevention. Data suggest t...

Clinical Trials [10797 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 ...

Use of Novel High-protein Food Products To Reduce Insulin Resistance

This controlled weight loss trial in adults at cardiometabolic risk (elevated triglyceride/HDL ratio) followed a randomized, cross-over design and utilized novel, high-protein pasta and ce...

Determination of the Postprandial Responses to Two Cereal Foods Differing by Their SDS Content Consumed Alone.

This study aimed at determining the 4-hours postprandial blood glucose and insulin responses to the ingestion of cereal product differing by their SDS content.

Glycemic and Insulinemic Impact of Oats With Additional Dried Fruits, Nuts and Seeds Soaked Overnight

The purpose of this study is to compare the glucose, insulin and subjective hunger responses elicited by oats with additional dried fruits, nuts and seeds soaked overnight in skim milk and...

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...

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).

Tools used in dentistry that operate at high rotation speeds.

Quick Search
Advertisement
 


DeepDyve research library

Relevant Topics

Food
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 ...

Nutrition
Within medicine, nutrition (the study of food and the effect of its components on the body) has many different roles. Appropriate nutrition can help prevent certain diseases, or treat others. In critically ill patients, artificial feeding by tubes need t...


Searches Linking to this Article