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Bloom Lab SARS-CoV-2 RBD SSM Libraries
(Pooled Libraries #1000000182, #1000000183, #1000000184, #1000000185, #1000000186, #1000000187, #1000000188)

  • Purpose

    Site-saturation mutagenesis (SSM) libraries of yeast-displayed SARS-CoV-2 receptor-binding domain (RBD). Each SSM library is in the background of fixed mutations that define certain SARS-CoV-2 variants that have evolved, including the ancestral Wuhan-Hu-1 strain and variants Alpha (N501Y), Beta (K417N/E484K/N501Y), Delta (L452R/T478K), Eta (E484K), and Omicron BA.1 (15 mutations) and BA.2 (16 mutations). Each library has a downstream N16 barcode that was previously linked to mutant identity via PacBio sequencing.

    Libraries are useful for deep mutational scanning characterization of the impacts of SARS-CoV-2 mutations on receptor- and antibody-binding.

  • Vector Backbone

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 1000000182 SARS-CoV-2 RBD Wuhan-Hu-1 SSM Library 1 $500 Add to Cart
Pooled Library 1000000183 SARS-CoV-2 RBD Alpha SSM Library 1 $500 Add to Cart
Pooled Library 1000000184 SARS-CoV-2 RBD Beta SSM Library 1 $500 Add to Cart
Pooled Library 1000000185 SARS-CoV-2 RBD Delta SSM Library 1 $500 Add to Cart
Pooled Library 1000000186 SARS-CoV-2 RBD Eta SSM Library 1 $500 Add to Cart
Pooled Library 1000000187 SARS-CoV-2 RBD Omicron BA.1 SSM Library 1 $500 Add to Cart
Pooled Library 1000000188 SARS-CoV-2 RBD Omicron BA.2 SSM Library 1 $500 Add to Cart
Available to Academic and Nonprofits Only

Library Details

  • Mutagenized CDS
    SARS-CoV-2 Receptor Binding Domain
  • Mutagenized region size
    603 bp

Each catalog item is a pair of duplicate libraries to be used separately as technical replicates, as described in (Starr and Greaney et al. 2020 Cell and Starr et al. 2022 bioRxiv).

Each catalog item also contains a separate bacterial stab of the parent control plasmid, as listed in the following table.

Pooled Library Background Parent Plasmid # Plasmids/library
1000000182 Wuhan-Hu-1 166782 ~50,000
1000000183 Alpha 184405 ~50,000
1000000184 Beta 184406 ~50,000
1000000185 Delta 184407 ~100,000
1000000186 Eta 184404 ~50,000
1000000187 Omicron BA.1 184408 ~100,000
1000000188 Omicron BA.2 184409 ~100,000
The parent plasmid is included with each library purchased. Please do NOT order this plasmid in addition to the library.

Library Shipping

Each library will be delivered as two pooled DNA libraries in microcentrifuge tubes on blue ice (tubes labeled lib1 and lib2). The tube contents will not necessarily be frozen. For best results, minimize freeze-thaws.

  • Volume
    ∼10 µL
  • Concentration
    25 ng/µL

You will also receive a bacterial stab of the parental-control plasmid. This plasmid is SHIPPED SEPARATELY at ambient temperature.

Depositor Comments

The Bloom Lab provides the following GitHub repositories:

The SARS-CoV-2 RBD Beta SSM Library is also described in Greaney et al. 2022 "A SARS-CoV-2 variant elicits an antibody response with a shifted immunodominance hierarchy" PLoS Pathog. doi: 10.1371/journal.ppat.1010248 (Link opens in a new window)

The SARS-CoV-2 RBD Delta SSM Library is also described in Greaney et al. 2022 "The SARS-CoV-2 Delta variant induces an antibody response largely focused on class 1 and 2 antibody epitopes" bioRxiv. doi: 10.1101/2022.03.12.484088 (Link opens in a new window)

How to cite this pooled library ( Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    SARS-CoV-2 RBD SSM Libraries were a gift from Jesse Bloom (Addgene # 100000018x)
  • For your References section:

    Shifting mutational constraints in the SARS-CoV-2 receptor-binding domain during viral evolution. Starr TN, Greaney AJ, Hannon WW, Loes AN, Hauser K, Dillen JR, Ferri E, Farrell AG, Dadonaite B, McCallum M, Matreyek KA, Corti D, Veesler D, Snell G, Bloom JD. bioRxiv. 2022.02.24.481899. doi: 10.1101/2022.02.24.481899.