A Comprehensive Overview of PCR Principles and Applications
In modern biology, Polymerase Chain Reaction (PCR) is an indispensable technology that allows for the amplification of DNA segments, thus increasing the quantity of DNA and providing a powerful tool for genome research and molecular diagnostics.
Principles of PCR
PCR operates on the basic principle of mimicking DNA replication in vitro using a polymerase enzyme, resulting in exponential amplification of DNA. The reaction requires a DNA template, primers, dNTPs (deoxynucleotide triphosphates), PCR buffer, and DNA polymerase, among other ingredients. PCR cycling conditions also play a crucial role in determining the experimental outcome.
PCR templates can be sourced from various DNA origins, such as genomic DNA (gDNA), complementary DNA (cDNA), and plasmid DNA. However, the composition or complexity of DNA can affect the optimal starting amount for PCR amplification. Optimization of the DNA starting amount is crucial to avoid risks such as nonspecific amplification due to high starting amounts or reduced yield due to low starting amounts.
Primers, synthetic DNA oligonucleotides typically containing 15-30 nucleotides, are designed to anneal to specific regions flanking the target sequence in the DNA template through sequence complementarity. Apart from sequence homology, the primer design must consider factors such as melting temperature (Tm), avoiding primer self-complementarity, and preventing primer-dimer formation to ensure specific amplification.
dNTPs are the building blocks for the new DNA strand synthesis during PCR. To prevent residual PCR product contamination, dUTP can be substituted for dTTP, coupled with uracil-DNA glycosylase (UDG) pretreatment, which excises uracil-containing DNA strands. This step effectively eliminates residual PCR products, reducing the risk of false-positive results.
PCR buffer provides an optimal chemical environment for DNA polymerase activity, typically maintaining a pH of 8.0-9.5. Magnesium ions (Mg2+) act as cofactors for DNA polymerase, facilitating dNTP binding and stabilizing the phosphate backbone's negative charge during primer-template complex formation.
DNA polymerases are essential components of PCR, synthesizing new complementary DNA strands from single-stranded DNA templates. Polymerases with proofreading activity ensure accurate DNA replication, crucial for applications such as molecular cloning and sequencing.
PCR cycling involves repeated denaturation, annealing, and extension steps to amplify target DNA sequences. Optimization of cycling conditions, including temperature and duration, is essential for efficient and specific amplification, considering factors such as template complexity and GC content.
PCR Applications
PCR finds widespread applications in genomic research, molecular diagnostics, genetic engineering, and population genetics. Common applications include DNA sequencing, genotyping, gene cloning, and pathogen detection, owing to its rapid, efficient, sensitive, and specific DNA amplification capabilities.
Hecin focuses on the diagnosis of respiratory system pathogens, and the Kits have the characteristics of high sensitivity and simple operation, are suitable for the rapid diagnosis of diseases caused by RSV infection, and are compatible with the fluorescent PCR platform. The product is made into a freeze-dried powder reagent, which has strong stability and can be transported and stored at room temperature, eliminating the trouble of low-temperature cold chain transportation and storage; the operation is simple and convenient, and the operator does not need complicated manual packaging operations, detectable.
Reagents for research use only (PCR-fluorescent probe method) | |
Item No. | Product name |
Respiratory virus | |
1 | 10 Multiple Respiratory Pathogens Nucleic Acid Test Kit (PCR-fluorescence probe method) |
2 | 14 Multiple Respiratory Pathogens Nucleic Acid Test Kit (PCR-fluorescence probe method) |
3 | 22 Multiple Respiratory Pathogens Nucleic Acid Test Kit (PCR-fluorescence probe method) |
4 | 24 Multiple Respiratory Pathogens Nucleic Acid Test Kit (PCR-fluorescence probe method) |
Influenza virus | |
5 | Avian Influenza Virus H10 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
6 | Avian Influenza Virus H5 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
7 | Avian Influenza Virus H5N1 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
8 | Avian Influenza Virus H5N6 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
9 | Avian Influenza Virus H6 Nucleic Acid Test Kit (PCR- fluorescence probe method) |
10 | Avian Influenza Virus H7 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
11 | Avian Influenza Virus H7N9 Nucleic Acid Test Kit (PCR- fluorescence probe method) |
12 | Avian Influenza Virus H9 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
13 | Avian Influenza Virus N1 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
14 | Avian Influenza Virus N19 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
15 | Avian Influenza Virus N2 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
16 | Avian Influenza Virus N6 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
17 | Avian Influenza Virus N9 Nucleic Acid Test Kit (PCR- fluorescence probe method) |
18 | Influenza A (H1N1) Virus (2009) Nucleic Acid Test Kit (fluorescent PCR method) |
19 | Influenza A Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
20 | Influenza A Virus Subtype H3N8 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
21 | Influenza A Virus Subtype H7N9 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
22 | Influenza A Virus Subtype H9N2 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
23 | Influenza A Virus Subtype H10N3 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
24 | Influenza A Virus Subtype H3N2 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
25 | Influenza A Virus Subtype H7 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
26 | Influenza A Virus Subtype H9 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
27 | Influenza B Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
28 | Seasonal Influenza Virus Subtype H3 Nucleic Acid Test Kit (fluorescent PCR method) |
29 | Influenza A Virus/Influenza B Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
30 | Influenza A Virus Subtype H1/H3 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
31 | IAV/IBV/ADV Nucleic Acid Test Kit(PCR-fluorescent probe method) |
32 | Avian Influenza Virus N1/N2/N6/N9 Nucleic Acid Test Kit (PCR- fluorescence probe method) |
33 | Avian Influenza Virus H5/H7/H9 Nucleic Acid Test Kit (PCR- fluorescence probe method) |
34 | Avian Influenza Virus H5/H7/H9/H10 Nucleic Acid Test Kit (PCR- fluorescence probe method) |
35 | 2019-nCoV/IAV/IBV Nucleic Acid Test Kit(PCR-fluorescent probe method) |
36 | Influenza B Virus Victoria / Yamagata Nucleic Acid Test Kit (PCR-fluorescent probe method) |
37 | pdmH1N1(HA)/H3N2(HA) Subtype Influenza Virus Nucleic Acid Test Kit (PCR- fluorescence probe method) |
38 | SARS-CoV-2/Flu/RSV Nucleic Acid Test Kit(PCR-fluorescence probe method) |
Adenovirus | |
39 | Adenovirus type 3/7 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
40 | Adenovirus type 21 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
41 | Adenovirus type 3 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
42 | Adenovirus type 4 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
43 | Adenovirus type 55 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
44 | Adenovirus type 7 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
45 | ADV Nucleic Acid Test Kit (PCR-fluorescent probe method) |
46 | Adenovirus type 11/14/55 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
47 | Adenovirus type 14 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
48 | Adenovirus type 16/34/50 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
49 | Adenovirus type 4/21/35 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
50 | Adenovirus type B/C/E Nucleic Acid Test Kit (PCR-fluorescence probe method) |
Coronavirus | |
51 | 2019-nCoV Nucleic Acid Test Kit(PCR-fluorescent probe method) |
52 | Coronavirus Nucleic Acid Test kit(PCR-fluorescent probe method) |
53 | HCoV 229E Nucleic Acid Test Kit (PCR-fluorescence probe method) |
54 | HCoV HKU1 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
55 | HCoV NL63 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
56 | HCoV OC43 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
57 | MERS coronavirus Nucleic Acid Test kit(PCR-fluorescent probe method) |
58 | Middle East respiratory syndrome coronavirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
59 | HCoV OC43／229E Nucleic Acid Test Kit (PCR-fluorescence probe method) |
60 | HCoV NL63／HKU1 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
Enterovirus | |
61 | CA16 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
62 | EV Nucleic Acid Test Kit (PCR-fluorescence probe method) |
63 | EV71 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
64 | Coxsackievirus 6/Coxsackievirus 10 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
65 | Enterovirus 71/Coxsackievirus A16/Enterovirus Universal Nucleic Acid Test Kit (PCR-fluorescent probe method) |
Rhinovirus | |
66 | Human Rhinovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
67 | Human Rhinovirus (Universal /C) Nucleic Acid Test Kit (PCR-fluorescence probe method) |
Parainfluenza virus | |
68 | PIV 1 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
69 | PIV 2 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
70 | PIV 3 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
71 | PIV 4 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
72 | Parainfluenza Virus Nucleic Acid Test Kit (PCR-fluorescent probe method) |
73 | Parainfluenza Virus 1/3 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
74 | Parainfluenza Virus 2/4 Nucleic Acid Test Kit (PCR-fluorescent probe method) |
Respiratory syncytial virus | |
75 | Respiratory Syncytial Virus type A Nucleic Acid Test Kit (PCR-fluorescence probe method) |
76 | Respiratory Syncytial Virus type B Nucleic Acid Test Kit (PCR-fluorescence probe method) |
77 | RSV Nucleic Acid Test Kit (PCR-fluorescent probe method) |
78 | Respiratory Syncytial Virus type A/B Nucleic Acid Test Kit (PCR-fluorescence probe method) |
Other microorganisms | |
79 | Acinetobacter Nucleic Acid Test Kit (PCR-fluorescence probe method) |
80 | B. Pertussis Nucleic Acid and Drug Resistance Mutation Site Detection Kit (PCR-fluorescent probe method) |
81 | Beta Hemolytic Streptococcus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
82 | Bordetella Pertussis Nucleic Acid Test Kit (PCR-fluorescence probe method) |
83 | Chlamydia Pneumoniae Nucleic Acid Test Kit (PCR-fluorescence probe method) |
84 | Clostridium Difficile Nucleic Acid Test Kit (PCR-fluorescence probe method) |
85 | Group A Streptococcus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
86 | Haemophilus Influenzae Nucleic Acid Test Kit (PCR-fluorescence probe method) |
87 | Klebsiella Pneumoniae Nucleic Acid Test Kit (PCR-fluorescence probe method) |
88 | Legionella Pneumophila Nucleic Acid Test Kit (PCR-fluorescence probe method) |
89 | Moraxella Catarrhalis Nucleic Acid Test Kit (PCR-fluorescence probe method) |
90 | MP Nucleic Acid Test Kit (PCR-fluorescence probe method) |
91 | Mycobacterium Tuberculosis Complex Nucleic Acid Test Kit (PCR-fluorescence probe method) |
92 | Mycobacterium Tuberculosis Ethambutol Resistance Mutation Nucleic Acid Test Kit (PCR-fluorescence probe method) |
93 | Mycobacterium Tuberculosis Fluoroquinolone Resistance Mutation Nucleic Acid Test Kit (PCR-fluorescence probe method) |
94 | Mycobacterium Tuberculosis Isoniazid Resistance Mutation Nucleic Acid Test Kit(PCR-fluorescence probe method) |
95 | Mycobacterium Tuberculosis Rifampicin Resistance Mutation Nucleic Acid Test Kit (PCR-fluorescence probe method) |
96 | Mycobacterium Tuberculosis Streptomycin Resistance Mutation Nucleic Acid Test Kit(PCR-fluorescence probe method) |
97 | Proteus Vulgaris Nucleic Acid Test Kit (PCR-fluorescence probe method) |
98 | Pseudomonas Aeruginosa Nucleic Acid Test Kit (PCR-fluorescence probe method) |
99 | Salmonella Enteritidis Nucleic Acid Test Kit (PCR-fluorescence probe method) |
100 | Shigella Flexneri Nucleic Acid Test Kit (PCR-fluorescence probe method) |
101 | Staphylococcus Aureus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
102 | Streptococcus Pneumoniae Nucleic Acid Test Kit (PCR-fluorescent probe method) |
103 | Vibrio Cholerae Nucleic Acid Test Kit (PCR-fluorescence probe method) |
104 | Vibrio Parahaemolyticus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
105 | Streptococcus Pneumoniae/Mycoplasma Pneumoniae/Haemophilus Influenzae Nucleic Acid Test Kit (PCR-fluorescent probe method) |
106 | Superbacteria (MRSA) Nucleic Acid Test Kit (PCR-fluorescence probe method) |
107 | Vibrio Cholerae General/01/0139/CTX gene Nucleic Acid Test Kit (PCR-fluorescence probe method) |
108 | Enteric Norovirus/Rotavirus/Adenovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
109 | Group A Streptococcus B/Streptococcus Pneumoniae Nucleic Acid Test Kit(PCR-fluorescence probe method) |
110 | Mycoplasma Pneumoniae/Chlamydia Pneumoniae /Legionella Pneumophila Nucleic Acid Test Kit (PCR-fluorescence probe method) |
111 | Mycoplasma Pneumoniae/Chlamydia Pneumoniae Nucleic Acid Test Kit (PCR-fluorescence probe method) |
Other virus | |
112 | African Swine Fever Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
113 | Astrovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
114 | Dengue Virus II/IV Nucleic Acid Test Kit (PCR-fluorescence probe method) |
115 | Dengue Virus I/III Nucleic Acid Test Kit (PCR-fluorescence probe method) |
116 | Dengue Virus Typing Nucleic Acid Test Kit (PCR-fluorescence probe method) |
117 | Echovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
118 | Echovirus Universal/11/30 Nucleic Acid Test Kit (PCR-fluorescence probe method) |
119 | Epstein-Barr Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
120 | HBoV Nucleic Acid Test Kit (PCR-fluorescent probe method) |
121 | Herpes Zoster Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
122 | Herps Simplex Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
123 | Human Cytomegalovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
124 | Human Metapneumovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
125 | Measles Virus Nucleic Acid Test Kit (PCR-fluorescent probe method) |
126 | Measles/Rubella Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
127 | Mpox Virus Nucleic Acid Test Kit (PCR- fluorescenceprobe method) |
128 | Mumps Virus Nucleic Acid Test Kit (PCR-fluorescent probe method) |
129 | Norovirus GⅡ Nucleic Acid Test Kit (PCR-fluorescence probe method) |
130 | Norovirus GI Nucleic Acid Test Kit (PCR-fluorescence probe method) |
131 | Norovirus GI/GⅡ/ Rotavirus A Nucleic Acid Test Kit (PCR-fluorescence probe method) |
132 | Rotavirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
133 | Rubella Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
134 | Sapovirus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
135 | Zika Virus Nucleic Acid Test Kit (PCR-fluorescence probe method) |
