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Optimization of Lyophilized LAMP and RT-PCR Reaction Mixes for Detection of Tuberculosis Cover

Optimization of Lyophilized LAMP and RT-PCR Reaction Mixes for Detection of Tuberculosis

The EuroBiotech Journal
Volume 4 (2020): Issue 4 (October 2020)
By: Esra Agel and  Hasan Sagcan  
Open Access
|Oct 2020

Figures & Tables

Figure 1

Results of different cryoprelyophilized mixture. 1) Freshly prepared mixture, 2) (0.5 % PEG 8000 + 2.0 % stachyose) Cryoprotectant mixture, 3) (0.5 % PEG 8000 + 1.0 % stachyose) Cryoprotectant mixture.
Results of different cryoprelyophilized mixture. 1) Freshly prepared mixture, 2) (0.5 % PEG 8000 + 2.0 % stachyose) Cryoprotectant mixture, 3) (0.5 % PEG 8000 + 1.0 % stachyose) Cryoprotectant mixture.

Figure 2

Results of lyophilized RT-PCR (a) and lyophilized LAMP (b) mixture. a: only 3. cryoprotectant mixture has positive result in RT-PCR assay. b: Results of LAMP assay 1) DNA ladder (100 bp, NEB), 2) 1. Cryoprotectant mixture, 3) 2. Cryoprotectant mixture, 4) 3. Cryoprotectant mixture 5) Freshly prepared mixture.
Results of lyophilized RT-PCR (a) and lyophilized LAMP (b) mixture. a: only 3. cryoprotectant mixture has positive result in RT-PCR assay. b: Results of LAMP assay 1) DNA ladder (100 bp, NEB), 2) 1. Cryoprotectant mixture, 3) 2. Cryoprotectant mixture, 4) 3. Cryoprotectant mixture 5) Freshly prepared mixture.

Figure 3

RT-PCR results of thermal stability of lyophilized mixtures
RT-PCR results of thermal stability of lyophilized mixtures

Figure 4

Detection limit results of lyophilized RT-PCR (a) and LAMP (b). a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control. b: a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control
Detection limit results of lyophilized RT-PCR (a) and LAMP (b). a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control. b: a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control

Figure 5

An image of lyophilized mixture in PCR tubes.
An image of lyophilized mixture in PCR tubes.

Cryoprotectant mixtures and amount of ingredients

The protocol of freeze dryer

DOI: https://doi.org/10.2478/ebtj-2020-0027 | Journal eISSN: 2564-615X
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Language: English
Page range: 230 - 236
Published on: Oct 21, 2020
Published by: European Biotechnology Thematic Network Association
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
Lyophilized master mix,
Loop-mediated isothermal amplification,
LAMP,
Real-time PCR
Related subjects:
Life sciences,
Genetics,
Biotechnology,
Bioinformatics,
Life sciences, other

© 2020 Esra Agel, Hasan Sagcan, published by European Biotechnology Thematic Network Association
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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