Experiment 4- Gene Expression Study in Arabidopsis Thaliana.

Experiment 4- Gene Expression Study in Arabidopsis Thaliana

Control

SRYTarget

RNA

RNA Blot

Seed mRNA Flower mRNA

Primers SpecificFor bobg mRNA

Quantitative RT-PCR

Microarrays

Tools For Studying Differential Gene Activity

What Are The Major Differences Between DNA and RNA?

RNA Has a Uracil Instead of Thymine

RNA Has Ribose Sugar in Nucleotide

Eukaryotic RNAs Are Single-Stranded Polynucleotides

NucleotidesJoined By

Phosphodiester Bonds Like All Nucleic Acids

Order 5’ 3’ Leads to Function:

Co-Linear With Gene Sequence

RNA Has Intra-Strand Double Helices or Secondary Structure

Total RNA Isolation Workflow

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Collecting plant

materials (1-6)

Lysis (7-11)

(12-16)

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Washing (21-32)

Precipitation & Binding

(17-20)

Elute RNA (33-

40)

Determining Concentratio

n

1. RNA Isolation

2. Remove Genomic DNA

RNase-Free DNase I

3. ElectrophoresisDetermining Quality

of Isolated RNA

Determining Quality of Isolated Total RNA Using A Capillary Gel Electrophoresis System

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Determining Quality of Isolated Total RNA Using A Capillary Gel Electrophoresis System

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Determining Quality of Isolated Total RNA Using A Capillary Gel Electrophoresis System-

Part II

Leaf

Leaf

Silique

Silique