Laboratory Report on Polymerase Chain Reaction (PCR) for Identifying Bacterial Colonies

Laboratory Report
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Table of Contents
AIM .................................................................................................................................................3
INTRODUCTION...........................................................................................................................3
MATERIALS...................................................................................................................................3
METHOD........................................................................................................................................4
For PCR..................................................................................................................................4
For agarose gel electrophoresis..............................................................................................5
For BLAST.............................................................................................................................6
DISCUSSION..................................................................................................................................6
CONCLUSION................................................................................................................................8
INTERPRETATION........................................................................................................................9
REFERENCES..............................................................................................................................10
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AIM
To identify the colonies from each plate and confirm their identity via using colony
Polymerase Chain Reaction with 16s rDNA.
INTRODUCTION
Polymerase Chain Reaction (PCR) has been considered as a general laboratory technique
which are commonly utilised for making billions of copies for a desired part of DNA. Specific or
universal primers are used for extending the desired fragment of gene in different cycles of
chain reaction (Huggett, Cowen and Foy, 2015). The prime goal of PCR is to form such a high
number of polymer which will be further employed in electrophoresis or sequencing. Three steps
are being implemented in PCR reaction which are denaturation, annealing and extension. Such
steps are accomplished at varying temperature depending upon the primers and template
sensitivity. The region may either code for any specific protein or may be required in forensic
studies to examine scene via DNA fingerprinting and sequencing of suspect. PCR is mainly
utilised in the fields like medical diagnosis, molecular research, biology, forensic or even in few
domains of ecology. Gel electrophoresis is the most popular technique which is employed in
separating the DNA fragments from each other on the basis of molecular size as well as charge,
where the heaviest particles being high in molecular weight remains near the well while smallest
molecule of DNA move farthest from the wells. The charge of such molecules are negative so
they tend to move towards the positive electrode. BLAST has referred to as an algorithm which
are being used for sequencing the DNA or proteins by comparing basic biological data of
nucleotide or amino acid sequence. The microorganism class and characteristic have been
determined by PCR followed by gel electrophoresis for separation and then sequencing NCBI
gene nucleotides with the help of BLAST (Priyam and et.al.,2019).
MATERIALS
Materials which are utilised in performing PCR, gel electrophoresis and BLAST are
mentioned below.
For PCR
 Bacterial colonies on plates which are labelled S1-S4
 Universal primer mix used for extending 16s rDNA sequence (forward and
reverse primer)
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