Running Header: lab report 1

Lab Report

Student’s Name

Professor’s Name

Course Title

Date
 Lab Report 2

DNA Extraction and Quantification – Quality Analysis

Introduction

In this experiment, the extraction and quantification analysis of genomic DNA will be done.

Consequently, both quality and quantity analysis of the same DNA will be done. Extraction of

DNA is separation, purification methods. Through this experiment, we learn how to obtain DNA

by destroying structures cell wall with different solutions. This experiment uses fractional by gel

electrophoresis method with agarose gel since proteins, essential oils, RNA are minimal at DNA

obtained from.

DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other

organisms. Nearly every cell in a person’s body has the same DNA. Most DNA is located in the

cell nucleus (where it is called nuclear DNA), but a small amount of DNA can also be found in

the mitochondria (where it is called mitochondrial DNA or mtDNA). DNA analysis techniques

are important in technologies such as genetic engineering, forensics, bioinformatics and in

anthropology.

Procedure

25µl of QIAGEN protease was pipetted into a 1.5 ml microcentrifuge tube, after which 200µl of

the sample (whole blood) was added. This was followed by the addition of 250µl of buffer P1.

the cap was closed and content mixed with pulse vortexing for 15 seconds. The sample was then

incubated at 56oc for 15 minutes in a heated block. The 1.5ml tube was then briefly centrifuged

to remove drops from inside the lid. 250µl of ethanol was added to the sample and mixed

thoroughly by pulse vortexing for 15 seconds. This was followed by five minutes incubation.

After another brief centrifugation, all the lysate was carefully applied into a QIAmp mini elute
 Lab Report 3

column without wetting the rim. This was followed by centrifugation at 13000rpm for 10

minutes. The column was then placed in a 2ml collection tube and the collection tube containing

the filtrate discarded. QIAmp mini elute column was carefully opened and 0.5 ml of buffer PB

was added without wetting the rim. After closing the cap, the sample was centrifuged at 13000

rpm for 1 minute. The elute column was again placed in a clean 2ml collection tube discarding

the filtrate. The elute column was carefully opened and 0.75 ml of buffer PB added. The tube

was centrifuged at 13000rpm for 1 minute. The elute column was placed in a new collection

tube, after which 500ml of ethanol was added and the sample centrifuged as before. The elute

column was then placed into a new clean 2ml collection tube and then centrifuged at full

speed(13000rpm) for 1 minutes to dry the membrane completely. After centrifugation, the elute

column was placed into a new column and the assembly incubated at 560c for 3 minutes to dry

the membrane. The elute column was finally placed in a 1.5ml microcentrifuge tube and the

collection tube with the filtrate discarded. Carefully, the lid was opened and 500µl of buffer PB

(RNAse free buffer) was applied at the center of the membrane, after which the lid was closed

and incubated at room temperature for 1 minute. This was followed by centrifugation at full

speed for 1 minute. The sample eluted in the 1.5 microcentrifuge tube contained the Nucleic

acids (DNA and RNA) of our interest.

Discussion

İt was possible to explain some few examples that needed to be considered when making the

genomic DNA extraction. For a prove for the correct experiment, air bubbles should not be

observed when adding dye and should not damage ultraviolet rays. The voltage applied to gel

should be watched with care not to dissolve the gel.When analyzed the image obtained as a result

we see that isolation is. Since its theoretically possible to degrade DNA which may not move and
 Lab Report 4

dissolve in the gel. Care is needed when performing experiment so that it has zero effect on the

gel which can move freely as opposed to DNA. İncase of no white line agoros gel which is

always obtained from DNA then it means DNA is present while the white spaces formed at the

bottom of the residue indicates the presence of RNAs.