Genomic and plasmid DNA isolations were performed with Bacterial & Yeast Genomic DNA Purification Kit (EURx Sp. z o.o., Gdańsk, Poland) and Plasmid Miniprep DNA Purification Kit (EURx Sp. z o.o., Gdańsk, Poland), respectively, according to the manufacturer’s protocols. Molecular cloning and transformation were performed according to standard protocols [70 ]. FastDigest restriction endonucleases were purchased in Thermo Fisher Scientific (Waltham, MA, USA). PCR was performed with high-fidelity Platinum SuperFi II DNA Polymerase (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s recommendations. The plasmids constructed and used in this work are listed in Supplementary Tables S2, S4, and S6–S8 . The primers used in this work were synthesized at Genomed S. A. (Warsaw, Poland) and are listed in Supplementary Tables S1, S2, and S5–S7 . Sanger DNA sequencing of plasmid constructs prepared in this work was performed in Genomed S. A. (Warsaw, Poland).
Bacterial yeast genomic dna purification kit
Manufactured by EURx
Sourced in Poland
The Bacterial & Yeast Genomic DNA Purification Kit is designed for the rapid and efficient extraction of high-quality genomic DNA from bacterial and yeast samples. The kit utilizes a silica-based membrane technology to provide a streamlined process for DNA purification.
Automatically generated - may contain errors
Lab products found in correlation
Plasmid miniprep dna purification kit, by EURx (2 mentions)
Phusion high fidelity dna polymerase, by Thermo Fisher Scientific (2 mentions)
Pcr dna clean up purification kit, by EURx (2 mentions)
Fastdigest restriction endonucleases, by Thermo Fisher Scientific (1 mentions)
Genejet plasmid miniprep kit, by Thermo Fisher Scientific (1 mentions)
Dreamtaq dna polymerase, by Thermo Fisher Scientific (1 mentions)
Oligonucleotide primer, by Merck Group (1 mentions)
Oligonucleotide primers, by Genomed (1 mentions)
Taq dna polymerase, by Thermo Fisher Scientific (1 mentions)
4 protocols using bacterial yeast genomic dna purification kit
1
Genomic and Plasmid DNA Isolation Protocol
2
Bacterial Genomic DNA Extraction and Sequencing
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Bacterial gDNA was extracted from cell pellets using a Bacterial & Yeast Genomic DNA Purification Kit (EURx, Poland) according to the manufacturer’s instructions. Based on the E. coli K-12 MG1655 genome sequence (RefSeq assembly accession: GCF_000005845.2) appropriate oligonucleotide primers were designed (Supplementary Table 2 ) for the amplification of fragments of the genes overlapping the targeted sequences. The PCR products were purified and sequenced.
3
Diverse DNA Manipulation Techniques
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Standard DNA manipulation methods including polymerase chain reactions (PCRs), restriction digests, ligations and DNA electrophoresis were performed as described previously [135 ]. Plasmid DNA was isolated using a GeneJET Plasmid Miniprep Kit (Thermo Scientific, Waltham, MA, USA). Genomic DNA was isolated using a Bacterial & Yeast Genomic DNA Purification Kit (EurX, Gdańsk, Poland). PCRs were performed with Phusion High-Fidelity DNA polymerase or DreamTaq DNA polymerase (Thermo Scientific). Oligonucleotide primers used for PCR were purchased from Sigma Aldrich and are listed in Supplementary Table S2 . DNA fragments amplified by PCR or obtained by restriction digestion were purified using a PCR/DNA Clean-Up Purification Kit (EurX). DNA sequencing was performed by Genomed S.A. (Warsaw, Poland).
4
Comprehensive DNA Manipulation Techniques
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All DNA manipulations, including the polymerase chain reaction (PCR), restriction digestions, ligations and DNA electrophoresis were performed as previously described (Sambrook and Russell, 2001) . Plasmid and chromosomal DNA were isolated using a Plasmid Miniprep DNA Purification Kit and Bacterial & Yeast Genomic DNA Purification Kit, respectively (EURx, Gdańsk, Poland). Restriction enzymes were obtained from Thermo Scientific (Waltham, USA). PCR was routinely performed in 25 µl or 50 µl reaction mixtures for 35 cycles using Taq DNA polymerase or, when fragments were used for cloning, Phusion High-Fidelity DNA Polymerase (Thermo Scientific). DNA fragments amplified by PCR were purified with a PCR/DNA Clean-Up Purification Kit (EURx) before and after restriction digestion. All kits and reagents were used according to the recommendations of the supplier.
Oligonucleotide primers used for PCR and sequencing were purchased from Genomed S.A.
(Warsaw, Poland) and are listed in Table S2. Plasmids used in this study are described in Table S1. DNA sequencing was performed by Genomed S.A. (Warsaw, Poland) .
Oligonucleotide primers used for PCR and sequencing were purchased from Genomed S.A.
(Warsaw, Poland) and are listed in Table S2. Plasmids used in this study are described in Table S1. DNA sequencing was performed by Genomed S.A. (Warsaw, Poland) .
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