a) What is the role of Trizol in RNA extractionTrizol reagent is .pdf

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a) What is the role of Trizol in RNA extraction? Trizol reagent is used for RNA isolation from cells and tissues. This reagent is used as it helps in maintaining RNA integrity during tissue homogenization, while at the same time disrupting and breaking down cells and cell components. The RNA remains only in the aqueous phase(Amrita). b) What was the purpose of performing a reverse transcription reaction. What were the products of thesereaction? Reverse transcription polymerase chain reaction (RT-PCR), a variant of polymerase chain reaction (PCR), is a technique commonly used in molecular biology to detect RNA expression incontrast the traditional PCR is used to exponentially amplify target DNA sequences, (Freeman, 1999). The product of this reaction will be RNA Solution a) What is the role of Trizol in RNA extraction? Trizol reagent is used for RNA isolation from cells and tissues. This reagent is used as it helps in maintaining RNA integrity during tissue homogenization, while at the same time disrupting and breaking down cells and cell components. The RNA remains only in the aqueous phase(Amrita). b) What was the purpose of performing a reverse transcription reaction. What were the products of thesereaction? Reverse transcription polymerase chain reaction (RT-PCR), a variant of polymerase chain reaction (PCR), is a technique commonly used in molecular biology to detect RNA expression incontrast the traditional PCR is used to exponentially amplify target DNA sequences, (Freeman, 1999). The product of this reaction will be RNA.

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• Define transcription • Define translation • What are the 3 steps of translation? • Define the “genetic dogma” • What is the function of Transfer RNA? • What is the function of RNA polymerase? • What is the function of DNA polymerase? • Define “splicing of RNA” • What is an exon? • What component of the cell does the translation? • What molecule in the cell does transcription? • What are the functions of: operon, promotor? • What is the difference between inducible operon and repressible operon? Solution • Define transcription Transcription is the process of making an RNA copy of a gene sequence. This copy, called a messenger RNA (mRNA) molecule, leaves the cell nucleus and enters the cytoplasm, where it directs the synthesis of the protein, which it encodes. Here is a more complete definition of transcription. • Define translation Translation is the process of translating the sequence of a messenger RNA (mRNA) molecule to a sequence of amino acids during protein synthesis. The genetic code describes the relationship between the sequence of base pairs in a gene and the corresponding amino acid sequence that it encodes. In the cell cytoplasm, the ribosome reads the sequence of the mRNA in groups of three bases to assemble the protein. Here is a more complete definition of translation: • What are the 3 steps of translation? Step # 1. Initiation: Initiation of translation in E .coli involves the small ribosome subunit, a mRNA molecule, a specific charge initiator tRNA, GTP, Mg++ and number of proteinaceous initiation factors (IFs). These are initially part of the small subunit and are required to enhance binding affinity of the various translational components (Table 8.1). Unlike ribosomal proteins, IFs are released from the ribosome once initiation is completed. Step # 2. Elongation: Once both subunits of the ribosome are assembled with the mRNA, binding site for two charged tRNA molecules are formed. These are designated as the ‘P’ or peptidyl and the ‘A’ or aminoacyl sites. The charged initiator tRNA binds to the P site, provided that the AUG triplet of mRNA is in the corresponding position of the small subunit. The increase of the growing polypeptide chain by one amino acid is called elongation. Step # 3. Termination: Termination of protein synthesis is carried out by triplet codes (UAG, UAA, UGA; stop codons) present at site A. These codons do not specify an amino acid, nor do they call for a tRNA in the A site. These codons are called stop codons, termination codons or nonsense codons. The finished polypeptide is still attached to the terminal tRNA at the P site, and the A site is empty. • Define the “genetic dogma” A theory in genetics and molecular biology subject to several exceptions that genetic information is coded in self-replicating DNA and undergoes unidirectional transfer to messenger RNAs in transcription which act as templates for protein synthesis in translation • What is the function of Transfer RNA? The tRNA molecule, or tr.

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