How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development

This survey aimed to assess the roles of pipette tip’s contamination and usage of one second-hand tip in the loading of all samples in the electrophoresis procedure, and use of second-hand tubes in PCR procedure. Three genes with different band sizes were amplified with the PCR method. Three dilutio...

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Main Authors: Elghar Soltani, Mohammad Ahangarzadeh Rezaee, Pourya Gholizadeh
Format: Article
Language:English
Published: Amber Publication 2019-04-01
Series:Journal of Research in Medical and Dental Science
Subjects:
PCR
Online Access:https://www.jrmds.in/articles/how-to-reuse-pipette-tips-and-tubes-in-pcr-and-electrophoresis-procedures-a-new-inhouse-method-development.pdf
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spelling doaj-6e62401f8f8e4617a95a7fd7b3cde9b72020-11-25T01:36:05ZengAmber PublicationJournal of Research in Medical and Dental Science2347-25452347-23672019-04-0172210213How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method DevelopmentElghar Soltani0Mohammad Ahangarzadeh Rezaee1Pourya Gholizadeh2Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, IranInfectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, IranStudent Research Committee, Tabriz University of Medical Sciences, Tabriz, IranThis survey aimed to assess the roles of pipette tip’s contamination and usage of one second-hand tip in the loading of all samples in the electrophoresis procedure, and use of second-hand tubes in PCR procedure. Three genes with different band sizes were amplified with the PCR method. Three dilutions of 1x, 0.5x and 0.1x were made for all three PCR products. After loading of each PCR products in its first well, pipette tip has not replaced and washed in 5 μl distilled water. Then, 5 μl of the washed water was loaded in the second well. Two kinds of staining methods were done, including safe stains and Ethidium- Bromide. At the same time, Non-sterile and second-hand vials were washed with 6% NaOCl and used for PCR method, again. Results showed that among all three dilutions (1x, 0.5x and 0.1x), only 1x dilutions had bands in first wells for each gene. In addition, none of them has presented bands in the second wells (which was done without replacing the pipette tips). In addition, 6% NaOCl had the ability to eliminate contamination from second-hand vials among PCR and vials could be reused for second times. We conclude that it is not necessary to replace the pipette tips among the loading of products in electrophoresis and second-hand tubes are usable after washing with NaOCl (6%-10%) for PCR.https://www.jrmds.in/articles/how-to-reuse-pipette-tips-and-tubes-in-pcr-and-electrophoresis-procedures-a-new-inhouse-method-development.pdfSecond-hand tubeReplacing of pipette tipElectrophoresisPCR
collection DOAJ
language English
format Article
sources DOAJ
author Elghar Soltani
Mohammad Ahangarzadeh Rezaee
Pourya Gholizadeh
spellingShingle Elghar Soltani
Mohammad Ahangarzadeh Rezaee
Pourya Gholizadeh
How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development
Journal of Research in Medical and Dental Science
Second-hand tube
Replacing of pipette tip
Electrophoresis
PCR
author_facet Elghar Soltani
Mohammad Ahangarzadeh Rezaee
Pourya Gholizadeh
author_sort Elghar Soltani
title How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development
title_short How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development
title_full How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development
title_fullStr How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development
title_full_unstemmed How to Reuse Pipette Tips and Tubes in PCR and Electrophoresis Procedures? A New In-house Method Development
title_sort how to reuse pipette tips and tubes in pcr and electrophoresis procedures? a new in-house method development
publisher Amber Publication
series Journal of Research in Medical and Dental Science
issn 2347-2545
2347-2367
publishDate 2019-04-01
description This survey aimed to assess the roles of pipette tip’s contamination and usage of one second-hand tip in the loading of all samples in the electrophoresis procedure, and use of second-hand tubes in PCR procedure. Three genes with different band sizes were amplified with the PCR method. Three dilutions of 1x, 0.5x and 0.1x were made for all three PCR products. After loading of each PCR products in its first well, pipette tip has not replaced and washed in 5 μl distilled water. Then, 5 μl of the washed water was loaded in the second well. Two kinds of staining methods were done, including safe stains and Ethidium- Bromide. At the same time, Non-sterile and second-hand vials were washed with 6% NaOCl and used for PCR method, again. Results showed that among all three dilutions (1x, 0.5x and 0.1x), only 1x dilutions had bands in first wells for each gene. In addition, none of them has presented bands in the second wells (which was done without replacing the pipette tips). In addition, 6% NaOCl had the ability to eliminate contamination from second-hand vials among PCR and vials could be reused for second times. We conclude that it is not necessary to replace the pipette tips among the loading of products in electrophoresis and second-hand tubes are usable after washing with NaOCl (6%-10%) for PCR.
topic Second-hand tube
Replacing of pipette tip
Electrophoresis
PCR
url https://www.jrmds.in/articles/how-to-reuse-pipette-tips-and-tubes-in-pcr-and-electrophoresis-procedures-a-new-inhouse-method-development.pdf
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