A polymerase chain reaction method for detecting dwarf mistletoe infection in Douglas-fir and western larch

1999 ◽  
Vol 29 (9) ◽  
pp. 1317-1321 ◽  
Author(s):  
M Marler ◽  
D Pedersen ◽  
T Mitchell-Olds ◽  
R M Callaway
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Method ◽  
Positive Control ◽  
Rbcl Gene ◽  
Larix Occidentalis ◽  
Chain Reaction ◽  
Dwarf Mistletoe ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Template Dna

Early detection and management of dwarf mistletoe (Arceuthobium spp.) is currently limited by the inability to rapidly detect infection during the 2- to 5-year endophyte phase of the parasite. We describe a polymerase chain reaction (PCR) technique for detecting Arceuthobium douglasii Engelm. and Arceuthobium laricis Engelm. in tissues of its hosts, Pseudotsuga menziesii (Mirb.) Franco and Larix occidentalis Nutt. DNA was extracted from branches of 15 infected and 15 uninfected P. menziesii. The PCR product amplified by using the Arceuthobium specific primer in the rbcL gene from Arceuthobium template DNA was a fragment of 708 pairs of bases in length. This product was amplified from all branches that were visibly infected, but the fragment was not generated from any samples known to be uninfected. The PCR product from conifer DNA was a fragment of 385 pairs of bases in length and was not amplified from pure mistletoe DNA; this was amplified as an internal positive control. The primers developed for P. menziesii and A. douglasii also worked on L. occidentalis and A. laricis. This method detected mistletoe DNA in 7 of 29 P. menziesii branches and 3 of 21 L. occidentalis branches that did not have external symptoms of infection and are presumed to be the result of the endophyte phase. This method provides a useful tool for experimental applications and for managing the spread of dwarf mistletoe.

scholarly journals Multiplex Polymerase Chain Reaction Method for Detection of Bovine Materials in Foodstuffs

2003 ◽  
Vol 86 (4) ◽  
pp. 764-767 ◽  
Author(s):  
Hong-Wei Gao ◽  
Da-Bing Zhang ◽  
Ai-Hu Pan ◽  
Wan-Qi Liang ◽  
Cheng-Zhu Liang
Keyword(s):  
Polymerase Chain Reaction ◽  
Genomic Dna ◽  
Polymerase Chain Reaction Method ◽  
Rapid Identification ◽  
Ribosomal Gene ◽  
Effective Control ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Pcr Product ◽  
Polymerase Chain

Abstract Rapid identification of bovine materials in animal foodstuffs is essential for effective control of a potential source of bovine spongiform encephalophathy. A convenient polymerase chain reaction (PCR)-based assay was developed for detection and identification of a bovine-specific genomic DNA sequence in foodstuffs. Simultaneously the assay assessed the DNA quality of the experiment system by amplification of a highly conserved eucaryotic DNA region of the 18-S ribosomal gene, helping to check the reliability of the test result. The amplified bovine-specific PCR product was a genomic DNA fragment of lactoferrin, a low copy gene that was different from a commonly used bovine-specific mitochondria sequence for identification of bovine materials. The specificity of this method was confirmed by the absence of detectable homologous PCR product using reference foodstuff samples that lacked bovine-derived meat and bonemeals, or genomic DNA samples from vertebrates whose offals are commonly included in animal feeds. This method could detect the presence of bovine material in foodstuffs when the samples contained >0.02% bovine-derived meat and bone meal. Furthermore, it was not affected by prolonged heat treatment. The specificity, convenience, and sensitivity of this method suggest that it can be used for the routine detection of bovine-derived materials.

scholarly journals Parallel DNA polymerase chain reaction: Synthesis of two different PCR products from a DNA template

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 320 ◽  
Author(s):  
Vikash Bhardwaj ◽  
Kulbhushan Sharma
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Polymerase ◽  
Chain Reaction ◽  
Single Stranded Dna ◽  
Pcr Product ◽  
Pcr Products ◽  
Parallel Direction ◽  
Polymerase Chain ◽  
Dna Template ◽  
Template Dna

Conventionally, in a polymerase chain reaction (PCR), oligonucleotide primers bind to the template DNA in an antiparallel complementary way and the template DNA is amplified as it is. Here we describe an approach in which the first primer binds in a parallel complementary orientation to the single-stranded DNA, leading to synthesis in a parallel direction. Further reactions happened in a conventional way leading to the synthesis of PCR product having polarity opposite to the template used. This is the first study showing that synthesis of DNA can happen also in a parallel direction. We report that from a single-stranded DNA template, two different but related PCR products can be synthesized.

scholarly journals A method for mapping germ line sequences in Tetrahymena thermophila using the polymerase chain reaction.

Genetics ◽  
1994 ◽  
Vol 137 (1) ◽  
pp. 95-106 ◽  
Author(s):  
D Cassidy-Hanley ◽  
M C Yao ◽  
P J Bruns
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Tetrahymena Thermophila ◽  
Germ Line ◽  
Repetitive Sequences ◽  
Mapping Technique ◽  
Ciliated Protozoan ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Template Dna

Abstract A method for mapping DNA sequences to specific germinal chromosomes in the ciliated protozoan Tetrahymena thermophila has been developed. This mapping technique (PCR mapping) utilizes the polymerase chain reaction and template DNA derived from nullisomic strains to directly assign micronuclear DNA sequences to specific micronuclear chromosomes. Using this technique, a number of unique sequences and short repetitive sequences flanked by unique sequences have been mapped to four of the five germinal chromosomes.

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