mlui hf (New England Biolabs)


Structured Review

Mlui Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mlui hf/product/New England Biolabs
Average 95 stars, based on 31 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Using a reverse genetics system to generate recombinant SARS-CoV-2 expressing robust levels of reporter genes"
Article Title: Using a reverse genetics system to generate recombinant SARS-CoV-2 expressing robust levels of reporter genes
Journal: bioRxiv
doi: 10.1101/2022.05.21.492922

Figure Legend Snippet: Assembly of entire SARS-CoV-2 genome into a BAC. a. The released five fragments (F1-F5) from pUC57 plasmids. b. Colony PCR screening for positive pBeloBAC-F1. c. Restriction digestion analysis of pBeloBAC-F13 using PacI and MluI. d. Restriction digestion analysis of pBeloBAC-F134 using PacI and MluI (F3); and MluI and BstBI (F4). e. Restriction digestion analysis of pBeloBAC-F1342 using KasI and PacI (F2); PacI and MluI (F3); and MluI and BstBI (F4). f. Restrict digestion confirmation of the pBeloBAC-FL using KasI and PacI (F2); PacI and MluI (F3); MluI and BstBI (F4); and BstBI and BamHI (F5).
Techniques Used: BAC Assay, Polymerase Chain Reaction

Figure Legend Snippet: Recovery of rSARS-CoV-2 from the pBeloBAC-FL. a. Protocol used for recovery of rSARS-CoV-2 from the pBeloBAC-FL using Lipofectamine 2000 transfection in Vero E6 cells. b. CPE caused by rSARS-CoV-2/WT in Vero E6 cells. Scale bars, 100 µm. c. Confirmation of the rescued rSARS-CoV-2/WT by IFA using a mouse antibody against viral N protein and a TRITC labeled donkey anti-mouse IgG secondary antibody. The nucleus was stained by DAPI. d. Sanger sequencing of the M gene of the natural isolate SARS-CoV-2 (top) and the recombinant SARS-CoV-2(bottom) that the MluI was removed by silent mutation.
Techniques Used: Transfection, Immunofluorescence, Labeling, Staining, Sequencing, Recombinant, Mutagenesis

Figure Legend Snippet: Overview of a BAC-based RG system for generation of rSARS-CoV-2. a. The schematic representation of the SARS-CoV-2 genome. Unique restriction sites used for vial genome assembly are indicated. b. Commercially synthesized five fragments (F1-F5) in pUC57 plasmids. Restriction sites used to release each of the viral fragments are indicated. The MluI and BstBI highlighted in red are the restriction sites that have been removed by silent mutation and used as genetics tags. c. Assembly of the entire SARS-CoV-2 genome into the empty pBeloBAC to generate the full-length rescue plasmid (pBeloBAC-FL).
Techniques Used: BAC Assay, Synthesized, Mutagenesis, Plasmid Preparation