Streptococcus agalactiae FPrA0245 (link) was kindly provided by Dr. Channarong Rodkhum of the Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok. E. coli DH10 beta, restriction enzymes, DNA ligase, DNA polymerase and dNTPs were products of New England Biolabs Inc. (England). Quick-change kit was purchased from Stratagene (USA). pET-28a was from Novagen (USA). Plasmid purification kit was from Geneaid (Taiwan). HisTrap FF affinity column was from GE Healthcare (England). Standard oligosaccharides (G1–G7) and LR-CD were products of Wako Pure Chemical Industry Ltd. and Ezaki Glico (Japan), respectively. Glucose oxidase kit was from Human (Germany). Pea starch (Emsland-Starke GmbH, Germany) was kindly provided by Prof. Wolfgang Zimmermann of the University of Leipzig, Germany. All chemicals used were of analytical grade.
E coli dh10 beta
Manufactured by New England Biolabs
Sourced in United Kingdom, United States
E. coli DH10 beta is a laboratory strain of Escherichia coli bacteria commonly used as a host for cloning and plasmid amplification. It is a chemically competent cell line that is designed for high-efficiency transformation and plasmid propagation.
Automatically generated - may contain errors
Lab products found in correlation
Restriction enzyme, by New England Biolabs (2 mentions)
T4 dna ligase, by Thermo Fisher Scientific (2 mentions)
Dna ligase, by New England Biolabs (1 mentions)
Dna polymerase, by New England Biolabs (1 mentions)
Histrap ff affinity column, by GE Healthcare (1 mentions)
Dntps, by New England Biolabs (1 mentions)
Gblock gene fragment, by Integrated DNA Technologies (1 mentions)
Gibson assembly, by New England Biolabs (1 mentions)
Gibson assembly kit, by New England Biolabs (1 mentions)
3 protocols using e coli dh10 beta
1
Recombinant FPrA02 Protein Production
2
Genetic Circuit Assembly and Induction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
New genetic parts were synthesized as gBlock gene fragments (Integrated DNA Technologies, CA) and assembled into full genetic circuit using Gibson assembly (New England Biolabs, MA, 2611L). The modified circuit was sequence verified and transformed into E. coli DH10‐beta (New England Biolabs, MA, C3019). Cultures were grown in culture tubes under different induction conditions, and RNA‐seq samples were prepared by following the protocols described above.
3
Genetic Engineering of Salmonella Mutants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
E. coli DH10-beta (New England Biolabs, USA) was used to clone and amplify plasmids. The plasmids were constructed using T4 DNA ligase (Thermo Fisher Scientific, USA) after digestion with restriction enzymes (New England Biolabs, USA) or using the Gibson assembly kit (New England Biolabs, USA). Speed pfu DNA polymerase (NanoHelix, Korea) was used for polymerase chain reaction (PCR) amplification. The primers used in the study were chemically synthesized by Macrogen, Korea and listed in Table S1.
The attenuated S. typhimurium strain ΔppGpp was deficient in ppGpp biosynthesis through disruption of relA and spoT genes [22] . S. typhimurium CNC018 (ΔppGpp ΔSPI-1 ΔSPI-2) was created from ΔppGpp bacterium via complete deletion of two Salmonella pathogenicity island gene clusters, SPI-1 and SPI-2, which are responsible for bacterial invasion into host cells (39 genes) and intracellular survival and replication (31 genes), respectively [39] [40] [41] . Bacterial transformation with plasmids was performed using an electroporator set at 2.
The attenuated S. typhimurium strain ΔppGpp was deficient in ppGpp biosynthesis through disruption of relA and spoT genes [22] . S. typhimurium CNC018 (ΔppGpp ΔSPI-1 ΔSPI-2) was created from ΔppGpp bacterium via complete deletion of two Salmonella pathogenicity island gene clusters, SPI-1 and SPI-2, which are responsible for bacterial invasion into host cells (39 genes) and intracellular survival and replication (31 genes), respectively [39] [40] [41] . Bacterial transformation with plasmids was performed using an electroporator set at 2.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!