scholarly journals Quantification of Fusobacterium nucleatum at Depths of Root Dentinal Tubules in the Tooth Using Real-time Polymerase Chain Reaction: An In Vitro Study

Cureus ◽  
2019 ◽  
Author(s):  
Arathi Ganesh ◽  
Aruna Kumari Veronica ◽  
Rupa Ashok ◽  
Preethi Varadan ◽  
Kandaswamy Deivanayagam
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
In Vitro Study ◽  
Fusobacterium Nucleatum ◽  
Vitro Study ◽  
Chain Reaction ◽  
Dentinal Tubules ◽  
Polymerase Chain

Quantitative expression analysis of blastocyst-derived gene transcripts in preimplantation developmental stages of in vitro-produced bovine embryos using real-time polymerase chain reaction technology

2004 ◽  
Vol 16 (8) ◽  
pp. 753 ◽  
Author(s):  
Nermin El-Halawany ◽  
Siriluck Ponsuksili ◽  
Klaus Wimmers ◽  
Markus Gilles ◽  
Dawit Tesfaye ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Expression Pattern ◽  
Embryo Development ◽  
Developmental Stages ◽  
Bovine Embryo ◽  
Chain Reaction ◽  
Quantitative Expression ◽  
Polymerase Chain

The main objective of the present study was to analyse the quantitative expression pattern of genes from a subtracted blastocyst transcriptome throughout the preimplantation developmental stages of in vitro-produced bovine oocytes and embryos. For this purpose, Day 5 morula (M) cDNAs were subtracted from Day 7 blastocyst (B) cDNAs (B–M) and used to establish a B–M subtracted cDNA library, as reported previously. From the total generated clones, 19 were analysed quantitatively. The mRNA samples isolated from pools of immature oocytes (n = 150), mature oocytes (n = 150) and two-cell (n = 80), four-cell (n = 40), eight-cell (n = 20), morula (n = 6) and blastocyst (n = 3) embryos were reverse transcribed and subjected to real-time polymerase chain reaction (PCR) using sequence-specific primers and SYBR green as the DNA dye. A relative standard curve method was used to analyse the real-time data taking the morula stage as a calibrator. Applying suppression subtractive hybridisation (SSH), a total of 71 clones, which represent 33 different expressed sequence tags, were generated and available for analysis. Most transcripts were analysed for the first time in bovine embryogenesis. The real-time PCR has validated the results of SSH positively for 84% (16/19) of transcripts, whereas 16% (3/19) showed deviation in the expression pattern from the one seen during SSH. Several transcript-specific expression patterns were observed for genes that play decisive roles in bovine embryogenesis. In addition to identification, accurately quantifying the expression profiles of transcripts during development will pave the way towards understanding the molecular mechanisms of embryogenesis and their potential role in early embryo development. Most importantly, the present study has contributed to the enrichment of bovine embryo gene collection by generating new transcripts involved in bovine embryo development.

scholarly journals Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study

2002 ◽  
Vol 35 (5) ◽  
pp. 487-490 ◽  
Author(s):  
Rozália F. Campos ◽  
Juracy B. Magalhães ◽  
Eliana A.G. Reis ◽  
Mitermayer G. Reis ◽  
Sonia G. Andrade
Keyword(s):  
Polymerase Chain Reaction ◽  
Trypanosoma Cruzi ◽  
In Vitro Study ◽  
High Sensitivity ◽  
Positive Control ◽  
Chain Reaction ◽  
Negative Results ◽  
Polymerase Chain ◽  
Positive Controls

To evaluate the sensitivity of polymerase chain reaction (PCR) to reveal known number of trypomastigote in the blood of mice, three separate experiments were done. First: To eight samples of 500mul of normal mice blood, one aliquot of 1, 2, 3, 4, 5, 10, and 50 trypomastigotes respectively, were added. Second and third: 10 aliquots with 1 and 10 with 2 trypomastigotes were added to samples of 500mul of normal mice blood. Positive control: 500mul of blood containing 100,000 trypomastigotes. For kDNA minicircles amplification by PCR the primers:S35 and S36 were used. PCR revealed products of 330 b.p in the positive controls. When only one sample with the aliquots of 1 or 2 trypomastigotes was examined, results were negative; results were positive with aliquots of 3 to 50 trypomastigotes. In the 2nd and 3rd experiments, 9/10 aliquots with one parasite and 9/10 with 2 trypomastigotes were positive revealing a high sensitivity of this reaction. In conclusion, the presence of one single parasite in 500mul of blood, is enough for a positive PCR. This method could be used as a complement to the various parasitological cure tests in treated mice, when low volumes of blood are individually examined.

scholarly journals In vitro characterization and analysis of probiotic species in the chicken intestine by real-time polymerase chain reaction

2019 ◽  
Vol 98 (11) ◽  
pp. 5840-5846 ◽  
Author(s):  
Revathi Shanmugasundaram ◽  
Mohamad Mortada ◽  
G Raj Murugesan ◽  
Ramesh K Selvaraj
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Chain Reaction ◽  
Chicken Intestine ◽  
In Vitro Characterization ◽  
Polymerase Chain
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