All chemicals were of reagent grade and purchased from commercial suppliers including Aladdin (Shanghai, China), Yuanye Bio-Technology (Shanghai, China), Sigma-Aldrich (St. Louis, MO, USA), unless otherwise noted. Phusion High-Fidelity DNA polymerase and restriction enzymes were purchased from Thermo-Fisher Scientific (Pittsburgh, PA, USA). T4 DNA ligase was purchased from New England Biolabs (Ipswich, MA, USA). DNA extraction kit and gel extraction kit were purchased from Promega (Madison, WI, USA). Oligonucleotide primers were ordered from GeneWiz (Suzhou, China). Ni2+-NTA resin was purchased from Genscript (Nanjing, China). Amicao® Ultra-15 filtration devices (molecular cutoff 10–100 KDa) were purchased from Millipore (Billerica, MA, USA). BCA protein assay kit and precast-gel for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were purchase from Solar Bio (Beijing, China).
Oligonucleotide primers
Manufactured by Genewiz
Sourced in China
Oligonucleotide primers are short, synthetic DNA sequences used in various molecular biology techniques. They serve as templates for the amplification or detection of specific DNA regions during processes such as PCR (Polymerase Chain Reaction).
Automatically generated - may contain errors
Lab products found in correlation
Gel extraction kit, by Promega (2 mentions)
Ni2 nta resin, by GenScript (2 mentions)
Precast gel for sodium dodecyl sulfate polyacrylamide gel electrophoresis, by Solarbio (2 mentions)
T4 dna ligase, by New England Biolabs (2 mentions)
Bca protein assay kit, by Solarbio (2 mentions)
Dna extraction kit, by Promega (2 mentions)
Phusion high fidelity dna polymerase, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride pvdf membrane, by Merck Group (1 mentions)
Goat anti human igg hrp, by Abcam (1 mentions)
6 protocols using oligonucleotide primers
1
Cloning and Purification of Recombinant Proteins
2
Specific Detection of Porphyromonas gingivalis
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The universal bacterial primer pair, 27F/1492R (forward, 5'-AGAGTTTGATCCTGGCTCAG-3' and reverse, 5'-ACGGCTACCTTGTTACGACTT-3') and the P. gingivalis specific primer pair, 404F/R (forward, 5'-AGGCAGCTTGCCATACTGCG-3' and reverse, 5'-ACTGTTAGCAACTACCGATGT-3'), were used as previously described (13-15 (link)). The primer pair 27F/1492R was used for the first round of PCR amplification, generating a full-length 16S rDNA product. P. gingivalis specific primers 404F/R, targeting the internal sequence of 16S rDNA, were used to detect P. gingivalis in the second round of PCR amplification. To increase sensitivity, nested PCR was performed based on the sequence of the 16S rRNA fragment of P. gingivalis genomic DNA. The expected size of the amplification product by the inner primer pair was 404 bp in length. Oligonucleotide primers were synthesized by Genewiz, Inc.
3
Constructing Deletion Mutants Using Lambda-Red
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Deletion mutants were constructed using the bacteriophage lambda-red recombinase system. All oligonucleotide primers used in this study were purchased from GENEWIZ (Nanjing, China). All mutants were created using the E. coli C600 strain, and homologous recombination constructions were generated using PCR-purified products harboring a selectable antibiotic resistance gene (ARG) and 60-nucleotide homology extensions. The ARG was removed by transforming the pCP20 plasmid carrying a flippase. Mutants were confirmed by PCR and sequencing. On the other hand, the C600 and mutants (adjust the OD600 to 1) were challenged to decimal dilutions of phage AH67C600_Q5 and AH67C600_Q9 ranging from 109 to 102, and finally perform phage plaque reading.
4
Oligonucleotide Primer Preparation and C. glutamicum Transformation
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Oligonucleotide primers used in this work were purchased from Genewiz (Suzhou, China) and listed in Table S1. All of the DNA manipulations were carried out following the standard procedure.
Transformation of C. glutamicum was performed as previously described (Ruan et al., 2015) .
Transformation of C. glutamicum was performed as previously described (Ruan et al., 2015) .
5
Glycosylation Knockout in Arabidopsis
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Col-0 A. thaliana (plant-specific glycosylation knockout mutant TKO Arabidopsis), pPhasBar plasmid, and EHA105 Agrobacterium tumefaciens strains were supplied by Jilin Agriculture University Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development. Escherichia coli DH5α was obtained from Genewiz Biotechnology (Suzhou, China). Restriction endonucleases HindIII, NcoI, and XbaI were purchased from New England Biolabs (Ipswich, MA, USA). Polyvinylidene fluoride (PVDF) membranes were purchased from Millipore. Goat anti-human IgG-HRP was purchased from Abcam (USA). Oligonucleotide primers were acquired from Genewiz Biotechnology (Suzhou, China). All chemicals were of the highest purity available.
6
Molecular Biology Reagents and Techniques
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All chemicals were of reagent grade and purchased from commercial suppliers including Aladdin (Shanghai, China), Yuanye Bio-Technology (Shanghai, China), Sigma-Aldrich (St. Louis, MO, USA), unless otherwise noted. Phusion High-Fidelity DNA polymerase and restriction enzymes were purchased from Thermo-Fisher Scienti c (Pittsburgh, PA, USA). T4 DNA ligase was purchased from New England Biolabs (Ipswich, MA, USA). DNA extraction kit and gel extraction kit were purchased from Promega (Madison, WI, USA). Oligonucleotide primers were ordered from GeneWiz (Suzhou, China). Ni 2+ -NTA resin was purchased from Genscript (Nanjing, China). Amicao ® Ultra-15 ltration devices (molecular cutoff 10-100 KDa) were purchased from Millipore (Billerica, MA, USA). BCA protein assay kit and precast-gel for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were purchase from Solar Bio (Beijing, China).
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