In this course you will learn:
In this course you will learn:
The basic concepts of the standard polymerase chain reaction (PCR) technique.
The criteria required to design a DNA primer for PCR.
The online programs we usually use to design a DNA primer.
Polymerase chain reaction (PCR)
You will learn in this section:
Steps involved in the PCR (PCR cycling): denaturation, annealing, extension
Components of the PCR reaction: DNA template, DNA polymerase, dNTPs, forward primer and reverse primer.
PCR amplification program.
Exponential amplification.
The size difference between the PCR amplification products of the first, second, and third cycle.
Criteria for PCR primer design
You will learn in this section a detailed explanation of the PCR primer design criteria and how they affect the primer sensitivity and stability including; primer length, primer melting temperature, primer annealing temperature, GC% of the primer, GC-clamp, cross homology and primer secondary structure.
Tools and methods
In this section you will learn how to:
Retrieve a Gene Sequence form NCBI, and Determine the Exact Location for Each Exon on the Chromosome Using Graphics.
Compare Different mRNA Transcripts and Select One to Evaluate a Gene Expression in Novel Cells.
Understand Primer3 Setting.
Calculate Primer Self-Complementarity Score.
Check for Primer Cross Homology Using BLAT.
In_silico PCR, How to confirm PCR amplification
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