High Throughput PAM Determination Assay Libraries
(Pooled Libraries #160132, #160133, #160134, #160135)
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Purpose
These pooled libraries encode CRISPR-Cas target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM). The libraries are used to determine the PAM requirements of Cas enzymes.
Each library differs in the CRISPR-Cas target site sequence, the orientation of the randomized nucleotide sequence, and the library complexity.
Libraries
- Library 160132 — p11-3’_8xN_PAM-site1 (RTW554)
- For characterizing Cas9 PAMs
- Randomized 8 nt PAM 3' of target site 1
- Library 160133 — p11-3’_8xN_PAM-site2 (RTW555)
- For characterizing Cas9 PAMs
- Randomized 8 nt PAM 3' of target site 2
- Library 160134 — p11-5’_10xN_PAM-site3 (RTW572)
- For characterizing Cas12a PAMs
- Randomized 10 nt PAM 5' of target site 3
- Library 160135 — p11-5’_10xN_PAM-site4 (RTW574)
- For characterizing Cas12a PAMs
- Randomized 10 nt PAM 5' of target site 4
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Vector Backbone
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
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Pooled Library | 160132 | p11-3'_8xN_PAM-site1 | 1 | $325 | Add to Cart | |
Pooled Library | 160133 | p11-3'_8xN_PAM-site2 | 1 | $325 | Add to Cart | |
Pooled Library | 160134 | p11-5'_10xN_PAM-site3 | 1 | Pending | ||
Pooled Library | 160135 | p11-5'_10xN_PAM-site4 | 1 | $325 | Add to Cart |
Library Details
Please see the Purpose section for library details.
Library Shipping
This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.
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Volume∼10 µL
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Concentration1 ng/µL
Resource Information
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Protocols
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Terms and Licenses
Academic/Nonprofit TermsIndustry Terms- Not Available to Industry
Trademarks- Zeocin® is an InvivoGen trademark.
Depositor Comments
The Kleinstiver Lab created a Github repository sharing the code to perform an analysis of data generated using the HT-PAMDA method.
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.
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For your Materials & Methods section:
High Throughput PAM Determination Assay library was a gift from Keith Joung and Benjamin Kleinstiver (Addgene #) -
For your References section:
Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Walton RT, Christie KA, Whittaker MN, Kleinstiver BP. Science. 2020 Apr 17;368(6488):290-296. doi: 10.1126/science.aba8853. Epub 2020 Mar 26. PubMed 32217751Scalable characterization of the PAM requirements of CRISPR-Cas enzymes using HT-PAMDA. Walton RT, Hsu JY, Joung JK, Kleinstiver BP. Nat Protoc. 2021 Feb 5. pii: 10.1038/s41596-020-00465-2. doi: 10.1038/s41596-020-00465-2. PubMed 33547443