Methodology Simplified preparation of a DNA ladder using PCR

Ty, Wang; Wang, L; Wang, F

doi:10.4238/vol10-3gmr1177

Cited by 3 publications

(1 citation statement)

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“…DNA ladder is one of the essential standard reagents in molecular biology laboratories (Cooney 1994 ; Parker et al 1977 ). DNA ladder could be made by restriction enzyme digestion, PCR amplification, or combination of both (Henrici et al 2017 ; Lan et al 2012 ; Murray and Monchawin 1994 ; Wang et al 2011 ). To utilize the isolated plasmid, seven pair of primers were designed to amplify different sizes of amplicon.…”

Section: Discussionmentioning

confidence: 99%

Homemade plasmid Miniprep solutions for affordable research in low-fund laboratories

et al. 2022

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As a consequence of Covid-19 pandemic, the basic lab consumables are in shortage, especially in the low-income countries. Thus, the main objective of the present study is to develop and evaluate homemade solution to isolate plasmid. To pursue this objective, RNase A was overexpressed in Bl21 DE3 cells (E. coli strain) and prepared as crude refolding reaction with proper activity. Also, lysis buffers, neutralization buffer, and washing buffers were prepared. The homemade miniprep kit showed successful isolation of the px48SpCas9 plasmid. The prepared plasmid purity was enough to be used successfully in PCR amplification. In addition, to get extra benefits from this study, seven primers were designed to match the plasmid backbone to produce DNA ladder (100–1500 bp). In conclusion, we were able to have attainable working solutions for plasmid miniprep and DNA ladder.

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