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[DOCX File]Western Connecticut State University
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The next step was to transform the insert containing plasmid into BL21 cells for protein expression. Isopropyl β-D-1-thiogalactopyranoside (IPTG) is necessary for protein expression. IPTG is similar to allolactose, which is needed to activate the lac operon. The difference between IPTG an allolactose is that IPTG can’t be broken down by the ...
bl21 competent cells protocol

[DOC File]Theranostics
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BL21 cells were collected, sonicated in cold PBS and purified with Glutathione S-transferase (GenScript, L00206) beads or Nickel-nitrilotriacetic acid (GenScript, L00250) beads according to the users’ manual. Protein extraction and immunoblotting
e.coli bl21

[DOC File]Protein Expression and Purification
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Competent BL21(DE3) cells are transformed with pET23-CBD-EGFP according to standard protocols {Sambrook, 1989 #5531}, and plated on LB-carb (100 g/ml carbenicillin) plates. We usually split the 200 l transformation volume over 2 plates. Plates are incubated at 37(C overnight. Day 2
e coli bl21 de3

[DOC File]Materials and Methods:
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Transformation into BL21 cells: Plasmids were transformed into chemically competent BL21 for high-level protein expression. Here, 8uL of plasmid was combined with 25uL competent cells. The reaction was incubated on ice for 30 min., heat shocked at 42°C for 40 seconds, placed on ice and to it, 80uL SOC media was added.
bl21 genotype

[DOCX File]mcr.aacrjournals.org
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Briefly, BL21 cells were transformed and grown in Luria broth (LB) to an OD 600 of ~0.8 at which point protein expression was induced by the addition of 250µM Isopropyl β-D-1-thiogalactopyranoside (IPTG). Cells were grown overnight at 16 ºC, pelleted, then resuspended into lysis buffer, 20 mM sodium phosphate (pH 8.0), 500 mM NaCl, 10 mM ...
bl21 strain

[DOC File]SUPPLEMENTAL METHODS - Harvard University
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His-ChmCter and His-ChmNter were expressed in BL21 cells and purified on Ni2+-NTA beads using manufacturer conditions (Qiagen). Different GST-fusions of c-Jun and MBF1 were expressed in DH5( and purified on Glutathione agarose beads according to manufacturer procedure (Invitrogen). Radiolabeled proteins were produced with the TNT T7 Quick ...
bl21 de3 transformation protocol

Competent Cells
Prepare an overnight streak plate of BL21 Comp Cells grown on an LB/KAN plate. Take Kanamycin out of freezer to thaw. Thoroughly clean (and bleach if necessary) 2 liter growth bottles and centrifuge bottles to be used for BL21 preparation. Label 1.5mL Eppendorf tubes and 15mL blue cap conical tubes with “BL-21” and today’s date.
bl21 de3

[DOCX File]Supplemental Figure 1 - Genes & Development
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May 09, 2014 · H-NS-H6 purification: BL21 (DE3) cells transformed with pLS015 were grown in 500 ml LB + 50 µg/ml kanamycin at 37°C to an OD600 of 0.6. Protein expression was induced by adding IPTG to 0.8 mM, and cells were grown at 30°C for 3 h.
bl21 de3 cells

Bl21 cells