Determining Annealing Temperatures for Polymerase Chain Reaction

  title={Determining Annealing Temperatures for Polymerase Chain Reaction},
  author={Angela R. Porta and Edward Enners},
ABSTRACT The polymerase chain reaction (PCR) is a common technique used in high school and undergraduate science teaching. Students often do not fully comprehend the underlying principles of the technique and how optimization of the protocol affects the outcome and analysis. In this molecular biology laboratory, students learn the steps of PCR with an emphasis on primer composition and annealing temperature, which they manipulate to test the effect on successful DNA amplification. Students… 

A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne

Three derived cleaved amplified polymorphic sequence (dCAPS) markers for detecting glyphosate resistance in Lolium perenne are designed and evaluated and showed that all three dCAPS markers could successfully detect mutations at codon-106 in the target enzyme.

Research on automatic preprocessing equipment of salivary analyte for PCR inspection of coronavirus.

Polymerase chain reaction (PCR) inspection of salivary analyte is performed by pretreatment, RNA extraction setup, RNA extraction, PCR setup, and the PCR process. However, the pretreatment process is

Polymerase Chain Reaction for Detection of Grasserie Virus, BmNPV in the Fifth Instar Larvae of Silkworm, Bombyx mori (L) (Race: PM x CSR 2 )

The mulberry silkworm, Bombyx mori L is a Lepidopteran insect, life cycle of which include: Egg, Larval Instars, Pupa within silky cocoon and adult moth, which make it a quite delicate venture, easily susceptible to viral and other diseases.

Detection of Grasserie Virus, BmNPV in the Fifth Instar Larvae of Silkworm, Bombyx mori (L) (Race: PM x CSR2) Through Polymerase Chain Reaction

Since 4,500 years, silkworm, Bombyx mori has become a purely domesticated insect, and like other domesticated animals, it is a quite delicate venture easily susceptible to a number of seasonal diseases.


Molecular techniques gave sensitive, accurate identification of infection, established prevalence in endemic areas and planning time for proper control strategy, and this gives the indication that the molecular methods were highly sensitive, specific and reliable methods for detecting Fasciola sp.

Identification of sex linked molecular markers in Indonesian giant freshwater prawn Macrobrachium rosenbergii

Male giant freshwater prawn grows faster than its female. Therefore, male mono sex culture is one of the solutions to improve aquaculture production. The all-male population of giant freshwater

Genetic diversity of shallots (Allium ascalonicum L.) from several locations in North Sumatra, Indonesia based on RAPD markers

Abstract. Hasanah Y, Mawarni L, Hanu, H, Lestami A. 2022. Genetic diversity of shallots (Allium ascalonicum L.) from several locations in North Sumatra, Indonesia based on RAPD markers. Biodiversitas



Random amplified polymorphic DNA PCR in the microbiology teaching laboratory: Identification of bacterial unknowns *

Random amplified polymorphic DNA (RAPD) analysis for the microbiology laboratory was adapted as a practical activity that allowed students to generate and analyze data for the identification of bacterial unknowns and gave students an appreciation of the value of applying a molecular biological method to a discipline‐specific question.

Aligning goals, assessments, and activities: an approach to teaching PCR and gel electrophoresis.

Developing and implementation of novel exercises that integrate the biological concepts of DNA structure and replication with the techniques of PCR and gel electrophoresis were found to be effective in enhancing student understanding of these concepts as shown by student performance across all learning goals.

General concepts for PCR primer design.

The purpose of this chapter is to explain basic rules of oligonucleotide primer design, and how the information deduced by primer design software can be rationally interpreted and manipulated to fit experimental needs.

Specificity, efficiency, and fidelity of PCR.

The purpose of current communication is to focus on the essential components of setting up an effective PCR, and discuss how each of these component may influence the specificity, efficiency, and fidelity of PCR.

The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on

The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years.

PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification

A total of 306 800 primers covering most known human and mouse genes can be accessed from the PrimerBank database, together with information on these primers such as Tm, location on the transcript and amplicon size.

Using Class Poster Sessions to Teach Intermediary Metabolism

In teaching a one-semester biochemistry course, it is found that using in-class poster sessions can greatly improve students' appreciation of metabolism and facilitate their mastery of both general principles and specific details.

Biochemistry. What poison? Bacterium uses arsenic to build DNA and other molecules.

Researchers describe GFAJ-1, a bacterial strain that they say can replace the phosphorus in its key biomolecules, including DNA, with the legendary poison arsenic, and speculate that organisms like GFAj-1 could have thrived in the arsenic-laden hydrothermal vent–like environments of early Earth.

Life without a wall or division machine in Bacillus subtilis

This work identifies a single point mutation that predisposes cells to grow without a wall in the highly tractable model bacterium Bacillus subtilis and shows that propagation of L-forms does not require the normal FtsZ-dependent division machine but occurs by a remarkable extrusion-resolution mechanism.

The Biology of Genomes, 5-9 May 2009, Cold Spring Harbor, New York. The bug and the bacterium: interdependent genomes.

The newly sequenced DNA of the pea Aphid reflects a long history of give-and-take between the genomes of the bug and a tiny bacterium called Buchnera aphidicola, scientists reported at the Biology of Genomes meeting.