La 320c

Manufactured by Eiken Chemical

Sourced in Japan

The LA-320C is a laboratory equipment designed for the measurement and analysis of various samples. It is a compact and versatile instrument that can be used for a wide range of applications in chemical, pharmaceutical, and materials science research and development.

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Lab products found in correlation

Loopamp fluorescent detection reagent, by Eiken Chemical (2 mentions) Loopamp dna amplification kit, by Eiken Chemical (1 mentions)

Close spelling variants of this product

LA-320C Realtime Turbidimeter Loopamp LA-320C Realtime Turbidimeter Loopamp Real-time Turbidimeter LA-320C Loopamp LA-320c Real-time turbidimeter LA-320C

6 protocols using la 320c

Loopamp™ Leishmania Detection Protocol

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Loopamp™ Leishmania Detection Kit was performed as per manufacturer’s instructions. Three μl extracted DNA was added to a Loopamp tube plus 27 μl DNA free water. This was run for 40 min at 65 °C and a final step of 80 °C for 2 min in a Loopamp LF-160 incubator in Suriname or a LA-320C in the Netherlands (both from Eiken Chemical Co., Japan). The final result of the Loopamp reaction was visualised via UV illumination and scored as being either positive; see Fig. 2 for examples. Loopamp reactions were performed and interpreted by personnel blinded from the results of the microscopy and CL Detect.Fig. 2

Typical examples of LAMP results. After the LAMP reaction, the samples are illuminated with UV light. A positive sample shows turbidity, a negative samples remains clear. C positive control sample: A and B positive samples; C and D negative samples

Schallig H.D., Hu R.V., Kent A.D., van Loenen M., Menting S., Picado A., Oosterling Z, & Cruz I. (2019). Evaluation of point of care tests for the diagnosis of cutaneous leishmaniasis in Suriname. BMC Infectious Diseases, 19, 25.

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Polymerase Spiral Reaction for Rapid Amplification

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Polymerase spiral reaction reactions were carried out at 65°C for 60 min in a final volume of 25 μl containing the following components: 8 U of Bst DNA polymerase; 12.5 μl buffer solution (pH 8.8, containing 20 mM (NH₄)₂SO₄, 100 mM KCl, 16 mM MgSO₄, 1.6 M betaine, 0.2% Tween 20, and 2.8 mM of each dNTP); and 2.0 μl DNA template. The amount of primers per reaction was 0.8 μM for IF and IB and 1.6 μM for Ft and Bt. Finally, the reaction mixture was covered with a protectant (patent: ZL201210371448.5, China) to prevent aerosol cross-contamination.
Amplification was monitored by two methods, turbidity monitoring with a real-time turbidimeter (LA-320c; Eiken Chemical, Japan) and direct visual in the presence of pH dye. A 1-μl volume of pH indicator (containing cresol red, 2.0 mM; phenol red, 0.6 mM) was added to the reaction tube; the color change from red to yellow for positive samples was visible by naked eye under natural light, while the negative reaction stayed red (Tanner et al., 2015 (link)). Each experiment was repeated at least three times.

Jiang X., Dong D., Bian L., Zou D., He X., Ao D., Yang Z., Huang S., Liu N., Liu W, & Huang L. (2016). Rapid Detection of Candida albicans by Polymerase Spiral Reaction Assay in Clinical Blood Samples. Frontiers in Microbiology, 7, 916.

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Optimizing MCDA Assay for Candida Detection

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In the MCDA reaction system, temperature was essential for influencing amplification efficiency. the amplification temperature of C. tropicalis-MCDA assay was optimized from 61 to 68°C with 1°C interval. Amplification mixtures with 1 μL of template of C. albicans were used as negative controls (NCs), and 1 μL of distilled water (DW) was used as a blank control. The MCDA amplicons were monitored using loopamp real-time turbidimeter LA-320C (Eiken Chemical Co., Ltd., Japan). Turbidity >0.1 was considered as positive result.

Wang Y., Zhao X., Cheng J., Tang X., Chen X., Yu H, & Li S. (2021). Development and Application of a Multiple Cross Displacement Amplification Combined With Nanoparticle-Based Lateral Flow Biosensor Assay to Detect Candida tropicalis. Frontiers in Microbiology, 12, 681488.

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LAMP Reaction Detection Methods

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The amplification products were detected using two methods; a real-time turbidimeter (La-320C; Eiken Chemical Co., Ltd.) at 650 nm and visual observation. Pyrophosphate ions are released during the LAMP reaction process and they form a white magnesium phosphate precipitate. This can be monitored using a real-time turbidimeter, drawing reaction curves every 6 sec with Mg²⁺ ions in the reaction buffer (13 (link)). For visual detection, 1 µl of Loopamp Fluorescent Detection reagent (Eiken Chemical Co., Ltd.), containing a metal indicator, was added into the reaction system prior to amplification. The reaction buffer initially turned orange because the calcein was quenched by Mn²⁺ ions. Then, during amplification the calcein was displaced by pyrophosphate ions from the calcein/Mn²⁺ complex, and the color changed from orange to green. By contrast, if no amplification occurred, no color change was observed.

Yu L., Li H., Zhao X., Wang X., Wei X., Lin W., Li P., Cui L, & Yuan J. (2017). Rapid visual detection of binary toxin producing Clostridium difficile by loop-mediated isothermal amplification. Experimental and Therapeutic Medicine, 14(5), 4781-4788.

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LAMP Assay for Vibrio Species

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In order to examine the availability of two LAMP primer sets, the LAMP amplification either for V. parahaemolyticus strains or V. vulnificus strains was carried out as the following description. Briefly, the traditional LAMP reaction was performed with the Loopamp Kit in a final volume of 25 μL containing 1.6 μM each FIP and BIP primers, 0.8 μM each LF and LB primers, 0.4 μM each F3 and B3 primers, 12.5 μL 2× reaction mix, 1 μL of Bst DNA polymerase (8 U), 1 μL FD and 1 μL DNA template.
The amplification mixtures of convention LAMP were heated at 62 °C for 1 h then at 85 °C for 5 min to stop the amplification. Three methods were employed to monitor the conventional LAMP reaction. First, a real-time turbidimeter (LA-320C, Eiken Chemical Co., Ltd., Tokyo, Japan) was used to minor the LAMP reaction by recording the optical density (OD) at 650 nm every 6 s. The turbidity readings were produced in real-time format and a turbidity threshold value of 0.1 was defined. Second, the positive reactions could be directly seen color change by FD reagent. Moreover, the amplification products were also detected by electrophoresis on 2% agarose gels with ethidium bromide staining. Mixture without DNA template was used as a negative control.

Wang Y., Li D., Wang Y., Li K, & Ye C. (2016). Rapid and Sensitive Detection of Vibrio parahaemolyticus and Vibrio vulnificus by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique. Molecules, 21(1), 111.

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LAMP DNA Amplification Protocol

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The LAMP method was performed using a Loopamp DNA amplification kit (Eiken Chemical, Tokyo, Japan). When it was necessary to confirm the amplification visually, the Loopamp fluorescent detection reagent (Eiken Chemical) was added to the reaction solution. The composition of the reaction solution was as follows: 10 ng of template DNA, 1 × reaction mix, 5 pmol of F3 primer, 5 pmol of B3 primer, 40 pmol of FIP primer, 40 pmol of BIP primer, 20 pmol of LF loop primer (optional), 20 pmol of LB loop primer (optional), 1 μL of fluorescent detection reagent (optional), and 1 μL of Bst DNA polymerase. The total volume was 25 μL. The amplification reaction was maintained at 63 °C for 1 h and then at 80 °C for 5 min to inactivate the enzyme. A turbidity measurement apparatus (LA-320C, Eiken Chemical) was used to confirm amplification in real-time. A fluorescent detection reagent was added to the reaction solution confirm amplification according to the presence or absence of fluorescence in the reaction solution after completion of the reaction under visible light and UV irradiation.

Sugano Y., Sakata K., Nakamura K., Hosokawa A., Kouguchi H., Suzuki T, & Kondo K. (2022). Loop-mediated isothermal amplification (LAMP) for rapid and easy identification of Omphalotus japonicus. Food Chemistry: Molecular Sciences, 5, 100115.

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