The P. aeruginosa strain PAO1 was supplied by the Laboratory Department of Tongji Medical Hospital (Huazhong University of Science and Technology, Wuhan, China). Specific pathogen-free (SPF) Ba1b/c mice (n = 60), six to eight-weeks old and weighting 20-30 g, were provided by the Experimental Animal Center in Tongji Medical College. The pGPU6/GFP/Neo- siRNA expression vector for short hairpin RNAs (shRNAs) was purchased from GenePharma (Shanghai, China). The restriction enzymes used for insertion preparation and validation of the shRNA fragments, BbsI, BamHI and PstI, were from Promega (Madison, WI, USA), and the T4 ligase was from Roche (Basel, Switzerland). The plasmid purification kit and reverse transcription (RT) PCR kit were from Promega. The PCR primers were synthesized by Shanghai Invitrogen Bio Co., Ltd. (Shanghai, China). The gel-recovery kit for purification of the final siRNA transfectable products was from TaKaRa (Shiga, Japan). The E-test strip came from AB Biodisk (Solna, Sweden). The antibodies against interleukin (IL-1b and IL-12 cytokines), secondary species-appropriate antibodies, and chemiluminescent etection reagent for use in Western blotting were from R&D Systems (Minneapolis, MN, USA).
Plasmid purification kit
Manufactured by Promega
Sourced in United States
The Plasmid Purification Kit is a laboratory tool designed to isolate and purify plasmid DNA from bacterial cultures. It enables the extraction and concentration of plasmid DNA for various downstream applications, such as cloning, sequencing, or transfection experiments.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using plasmid purification kit
1
Establishment of P. aeruginosa Infection Model
2
Isolation and Manipulation of Bacterial Strains
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The bacterial strains and plasmids are listed in Table 1. S. aureus strains were cultured at 37°C in Tryptic Soy Broth (TSB; Oxoid, Basingstoke, UK) or on Tryptic Soy Agar (Oxoid); when necessary, the media were supplemented with either chloramphenicol (10 μg/ml) or anhydrotetracycline hydrochloride (50 ng/ml). Escherichia coli strains were grown at 37°C in Luria-Bertani liquid medium or on Luria-Bertani agar containing a suitable antibiotic: ampicillin or kanamycin (50 μg/ml), as appropriate.
Genomic DNA was extracted by TIANamp Bacterial DNA Kit (Tiangen Biotech Co., Ltd., Beijing, China). Plasmids from E. coli were extracted using a plasmid purification kit (Promega, WI, USA); for plasmid extraction of S. aureus, the cells were predigested with a digestion buffer with 40 U/ml lysostaphin, 10 mg/ml lysozyme and 10% (v/v) glycerol.
Genomic DNA was extracted by TIANamp Bacterial DNA Kit (Tiangen Biotech Co., Ltd., Beijing, China). Plasmids from E. coli were extracted using a plasmid purification kit (Promega, WI, USA); for plasmid extraction of S. aureus, the cells were predigested with a digestion buffer with 40 U/ml lysostaphin, 10 mg/ml lysozyme and 10% (v/v) glycerol.
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