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Experiment 4

This document provides information about a lab experiment on isolating and characterizing nucleic acids (DNA and RNA). The document includes pre-lab questions to help prepare students, data sheets to record results, and post-lab questions to analyze the experiment. Some key details are: - The pre-lab for DNA discusses differences between DNA and RNA structures, prokaryotic and eukaryotic DNA, and the expected A260/A280 ratio range for pure DNA. - The DNA data sheets are blank and left to be filled in during the experiment. - The post-lab asks students to explain reagents used in different DNA isolation methods and chemical tests used to characterize DNA. - The RNA

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Jio Santos
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351 views

Experiment 4

This document provides information about a lab experiment on isolating and characterizing nucleic acids (DNA and RNA). The document includes pre-lab questions to help prepare students, data sheets to record results, and post-lab questions to analyze the experiment. Some key details are: - The pre-lab for DNA discusses differences between DNA and RNA structures, prokaryotic and eukaryotic DNA, and the expected A260/A280 ratio range for pure DNA. - The DNA data sheets are blank and left to be filled in during the experiment. - The post-lab asks students to explain reagents used in different DNA isolation methods and chemical tests used to characterize DNA. - The RNA

Uploaded by

Jio Santos
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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PRE-LAB

Isolation and Characterization of Nucleic Acids (DNA)

Name: ​______________________________________​ Group No:_​3​____________


Section: ​2C-MT​______________________________ Date Submitted: ​________

1. State three major differences between the structures of DNA and RNA.
- DNA bases are A, G, C, and T; TNA bases are A, G, C, and U
- The sugar in DNA is 2-deoxy-D-ribose; in RNA it is D-ribose
- DNA is always double stranded; there are several kinds of RNA, all of which
are single stranded

2. Give at least two differences between prokaryotic and eukaryotic DNAs.


Prokaryotic DNA can be found in the cytoplasm while eukaryotic DNA is found in
the nucleus of the cell. Prokaryotic DNA also contains less genes, has no
histones and their DNA replication is faster than eukaryotic DNA and vice versa.
3. What is the range of expected A260/A280 ratio for pure DNA isolates?
DATA SHEETS
Isolation and Characterization of Nucleic Acids (DNA)

Isolation of DNA and UV measurement


Concentration
DNA Physical UV Measurement
Description (A​260​/A​280​) Protein Nucleic Acid
(mg/mL) (ug/mL)

Bacterial DNA White - - -


mucus-like
substance

Plant DNA Thread-like/ - - -


fibrous

Animal DNA White - - -


string-like
strands

Chemical Characterization
Chemical test Bacterial DNA Plant DNA Onion DNA Animal DNA
CTAB Mtd

Test for Upper layer:


Deoxyribose Dark brown

Lower layer:
clear/colorless

Test for Clear/colorless


Phosphate with yellow ppt.

Test for White cloudy


Pyrimidines solution
POST-LAB
Isolation and Characterization of Nucleic Acids (DNA)

1. Explain why the following reagents are used in the isolation of DNA: (meanne)
a. Microbial DNA Isolation
i. EDTA

Ii. Lysozyme

b. Plant DNA Isolation (CTAB Method)


i. CTAB

Ii. NaCl

c. DNA Isolation from an Onion


i. SDS

ii. Papain or meat tenderizer

d. Animal DNA
I. CHCl3: Isoamyl Alcohol

Ii. Isopropyl Alcohol

2. Explain the principle involved in the following chemical tests for the
characterization of DNA:
a. Test for deoxyribose

b. Test for phosphate

c. Test for purines

d. Test for pyrimidines


PRE-LAB
Isolation and Characterization of Nucleic Acids (RNA)

1. Identify the major types of RNA in a cell and discuss their individual functions in
relation to protein synthesis.

2. Compare the structural differences of sugars and bases in DNA and RNA
molecules.

3. What processing events differentiate eukaryotic mRNA from prokaryotic mRNA?


DATA SHEETS
Isolation and Characterization of Nucleic Acids (RNA)

Isolation of RNA and UV Measurement


RNA Physical UV Measurement Total RNA (J19)
Description
( A260 /A280 ) ( A260 /A230 )

Yeast White sediment - - -

Young Plant - - -

Chemical Characterization
Chemical Test RNA from Yeasts RNA from a Young Plant

Test for Ribose Green solution

Test for Purines Yellowish brown ppt

Test for Pyrimidines Brown ppt

Litmus: Red to Blue


POST-LAB
Isolation and Characterization of Nucleic Acids (RNA)

1. Explain why the following reagents are used in the isolation of RNA: (meanne)
a. Guanidine isothiocyanate

b. B-mercaptoethanol

c. Phenol

d. CH 3 COON a

2. Why is RNA more susceptible to alkaline hydrolysis than DNA?


(meanne)

3. Provide the chemical nature or composition of the compound formed in a positive


result for the following:
a. Test for ribose

b. Test for phosphate

c. Test for purines

d. Test for pyrimidines

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