Start Validating internal controls for quantitative plant gene expression studies

Validating internal controls for quantitative plant gene expression studies

Although this technique is still used to assess gene expression, it requires relatively large amounts of RNA and provides only qualitative or semi quantitative information of m RNA levels.

This measurement is made after each amplification cycle, and this is the reason why this method is called real time PCR (that is, immediate or simultaneous PCR).

In the case of RNA quantitation, the template is complementary DNA (c DNA), which is obtained by reverse transcription of ribonucleic acid (RNA).

In this instance the technique used is quantitative RT-PCR or Q-RT-PCR.

For this reason a number of standardization systems (often called normalization methods) have been developed.

Some have been developed for quantifying total gene expression, but the most common are aimed at quantifying the specific gene being studied in relation to another gene called a normalizing gene, which is selected for its almost constant level of expression.

(2002) Physiological responses of vascular plants to heavy metals.

In ‘Physiology and biochemistry of metal toxicity and tolerance in plants’.

In order to amplify small amounts of DNA, the same methodology is used as in conventional PCR using a DNA template, at least one pair of specific primers, deoxyribonucleotides, a suitable buffer solution and a thermo-stable DNA polymerase.