Title
Occurrence of microRNA expressions using two macronutrient deficiencies presented to Arabidopsis Thaliana
Introduction
Arabidopsis Thaliana was the model chosen for this experiment. It was used for miRNA expression because its entire genome is already sequenced (Weems 362-369). It is also easy to grow in difficult conditions and has a short lifespan. This all makes it an easy plant to work with for research (Weems 362-369). The macronutrients phosphorus and sulfur were used for the study because they are essential for plant growth (Axtell). Both phosphorus and sulfur are components of coenzymes and proteins (Axtell). Controlling gene expression via miRNA is the plant’s way of responding to limiting levels of these essential macronutrients. miRNA works by binding to single stranded mRNA which creates double stranded RNA. Thus the plant identifies the new double stranded mRNA as inferior and attacks it.
For the experiment, there were four microRNAs used including miR156, miR395, miR398, and miR399. miR156 was the control and has no nutrient requirement so it would be expected to not show any changes in nutrient levels …show more content…
Then the miRNAs would be extracted from each of the three mediums using a Sigma mirPremier miRNA isolation kit and then analyzed using qt-PCR. The control used was the full media plate. The media type that the seeds were grown on was the independent variable in the experiment while the dependent variable was the different levels of miRNA expression. The plants were then collected into a tube and grinded in a lysis mix with a pestle. To isolate the small RNAs, the Sigma mirPremier miRNA isolation kit was used. Reverse Transcriptase Master Mix was then used for the reverse transcription reactions by putting the miRNAs in a thermocycler to convert them into single stranded DNA. Finally, qt-PCR was used to analyze the levels of
Occurrence of microRNA expressions using two macronutrient deficiencies presented to Arabidopsis Thaliana
Introduction
Arabidopsis Thaliana was the model chosen for this experiment. It was used for miRNA expression because its entire genome is already sequenced (Weems 362-369). It is also easy to grow in difficult conditions and has a short lifespan. This all makes it an easy plant to work with for research (Weems 362-369). The macronutrients phosphorus and sulfur were used for the study because they are essential for plant growth (Axtell). Both phosphorus and sulfur are components of coenzymes and proteins (Axtell). Controlling gene expression via miRNA is the plant’s way of responding to limiting levels of these essential macronutrients. miRNA works by binding to single stranded mRNA which creates double stranded RNA. Thus the plant identifies the new double stranded mRNA as inferior and attacks it.
For the experiment, there were four microRNAs used including miR156, miR395, miR398, and miR399. miR156 was the control and has no nutrient requirement so it would be expected to not show any changes in nutrient levels …show more content…
Then the miRNAs would be extracted from each of the three mediums using a Sigma mirPremier miRNA isolation kit and then analyzed using qt-PCR. The control used was the full media plate. The media type that the seeds were grown on was the independent variable in the experiment while the dependent variable was the different levels of miRNA expression. The plants were then collected into a tube and grinded in a lysis mix with a pestle. To isolate the small RNAs, the Sigma mirPremier miRNA isolation kit was used. Reverse Transcriptase Master Mix was then used for the reverse transcription reactions by putting the miRNAs in a thermocycler to convert them into single stranded DNA. Finally, qt-PCR was used to analyze the levels of