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DNA primer design, what your mentor never taught you
An excellent training about Science
Primer Design For Polymerase Chain Reaction-Tips & Tricks
In this course you will learn: The basic concepts of the standardpolymerase chain reaction (PCR) technique. The criteria required to design aDNA primer for PCR. The onlineprograms we usually useto design a DNA primer. Tips for troubleshooting gel electrophoresis results. Polymerase chain reaction (PCR) You will learn in this section: Steps involved in the PCR (PCR cycling): denaturation, annealing, extensionComponents 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 designYou will learn in this section a detailed explanation of thePCR primer design criteria and how they affect the primer sensitivity andstabilityincluding; primer length, primermelting temperature, primerannealing temperature, GC% of the primer, GC-clamp, cross homology and primer secondary structure. Tools and methodsIn 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. Evaluate Primers Depending on Delta G.Gel Electrophoresis Troubleshooting In this section you will findtips for successful gelelectrophoresis. It includesall the possible problems you may encounter, and suggest you a solution for each problem.
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