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CRISPR header icon

Multiplex CRISPR/Cas9 Assembly System Kit
(Kit # 1000000055 )

Depositing Lab: Takashi Yamamoto

This 18 plasmid CRISPR kit is used to construct all-in-one CRISPR/Cas9 vectors expressing multiple gRNAs with either Cas9 nuclease or Cas9 D10A nickase. A separate accessory pack is available for dCas9 and FokI-dCas9.

This kit consists of one 96-well plate, and will be shipped as bacterial glycerol stocks on dry ice. Samples should be frozen at -80°C immediately upon arrival. Individual plasmids can be ordered at $65 per plasmid, and will be shipped as bacterial stabs

Add to Cart
$330 USD + shipping

Available to academics and nonprofits only.

Original Publication

Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400. PubMed .

Description

The Multiplex CRISPR/Cas9 Assembly System Kit enables construction of all-in-one CRISPR/Cas9 vectors expressing multiple gRNAs (2-7) and a Cas9 nuclease or nickase . Multiple gRNA cassettes are assembled using BsaI-mediated Golden Gate cloning method. The kit contains 18 plasmids: pX330A-1x[2-7], pX330A_D10A-1x[2-7], and pX330S-[2-7].

Overview of Multiplex CRISPR/Cas9 assembly

Illustration of CRISPR all-in-one vector assembly

Figure from Sakuma et al., Sci Rep . 2014 Jun 23;4:5400.

Kit Documentation

Icon Multiplex CRISPR-Cas9 Assembly System Kit.pdf (262.7 KB)

Protocol

Download a PDF Protocol for Multiplex CRISPR construction as described in the above article.

Icon Multiplex CRISPR Kit assembly protocol (365.9 KB)

Resources

Download a ZIP file containing all Multiplex CRISPR Kit plasmid sequences in GenBank format

Icon Multiplex CRISPR Kit plasmids - GenBank files (126.1 KB)

How to Cite this Kit

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

For your Materials and Methods section:
"The CRISPR kit used for constructing multiplex CRISPR/Cas9 vectors was a gift from Takashi Yamamoto (Addgene kit # 1000000054)"

For your Reference section:
Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400. PubMed .

Multiplex CRISPR/Cas9 Assembly System Kit - #1000000055

Resistance Color Key

Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.

Inventory

Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right. The Plasmid column links to a plasmid's individual web page.

Kit Plate Map

96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.

Resistance Color Key

Ampicillin
Spectinomycin

Inventory

Well Plasmid Resistance
A / 1 pX330A-1x2
Ampicillin
A / 2 pX330A-1x3
Ampicillin
A / 3 pX330A-1x4
Ampicillin
A / 4 pX330A-1x5
Ampicillin
A / 5 pX330A-1x6
Ampicillin
A / 6 pX330A-1x7
Ampicillin
A / 7 pX330A_D10A-1x2
Ampicillin
A / 8 pX330A_D10A-1x3
Ampicillin
A / 9 pX330A_D10A-1x4
Ampicillin
A / 10 pX330A_D10A-1x5
Ampicillin
A / 11 pX330A_D10A-1x6
Ampicillin
A / 12 pX330A_D10A-1x7
Ampicillin
B / 1 pX330S-2
Spectinomycin
B / 2 pX330S-3
Spectinomycin
B / 3 pX330S-4
Spectinomycin
B / 4 pX330S-5
Spectinomycin
B / 5 pX330S-6
Spectinomycin
B / 6 pX330S-7
Spectinomycin
Data cached @ 2017-08-19

Kit Plate Map - #1000000055

Plate #1

  1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
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