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MetClo DNA Assembly Kit
(Kit # 1000000153 )

Depositing Lab:   Christopher A. O'Callaghan

The MetClo DNA Assembly Kit is a 48 vector set allowing modular, hierarchical assembly of large DNA fragments using a single type IIS restriction enzyme. The kit includes the 3 E. coli strains necessary for BsaI, BpiI and LguI-based MetClo assembly vector preparation.

This kit will be sent as bacterial glycerol stocks in 96-well plate format.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

Original Publication

MetClo: methylase-assisted hierarchical DNA assembly using a single type IIS restriction enzyme Lin D, O'Callaghan CA. Nucleic Acids Research. 2018 Jul 9. pii: 5050631. doi: 10.1093/nar/gky596. PubMed PMID: 29986052.

Description

MetClo allows fast and flexible modular assembly of large DNA fragments from multiple donor plasmids using a single type IIS restriction enzyme. It relies on methylation-switching to selectively block an outside flanking pair of restriction sites but allows cutting at the inner flanking sites to generate compatible adhesive ends. Any fragments that start with adhesive end ‘p’ and end with adhesive end ‘q’ can then be cloned into any of the assembly vectors with one of three types of adaptor sequence (types ‘Start’, ‘Middle’, ‘End’). This allows insert fragments to be preferentially assembled in a one-pot reaction.


MetClo approach for rapid hierarchical assembly of large DNA fragments

Figure Legend: The standard MetClo vector set contains 30 low copy vectors with a p15a replication origin and either an ampicillin (Amp), kanamycin (Kan) or chloramphenicol (Cam) selection marker, and a LacZa negative selection marker flanked by different adaptor sequences sufficient for the assembly of up to 6 fragments in a single reaction. The vector set also contains 18 single copy vectors with an F replication origin, one of the three antibiotic selection markers, and a fragment containing a ColE1 replication origin and a LacZα negative selection marker flanked by different adaptor sequences sufficient for assembly of up to 4 fragments in a single reaction.

Kit-Specific Resources

Click below to download the MetClo Strain Verification protocol:

Icon MetClo Strain Verification protocol (66.3 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 MetClo DNA Assembly Kit was a gift from Christopher A. O'Callaghan (Addgene kit #1000000153).”

For your Reference section:

MetClo: methylase-assisted hierarchical DNA assembly using a single type IIS restriction enzyme Lin D, O'Callaghan CA. Nucleic Acids Research. 2018 Jul 9. pii: 5050631. doi: 10.1093/nar/gky596. PubMed PMID: 29986052.

MetClo DNA Assembly Kit - #1000000153

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 lists the plasmid well location in its plate. 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

Chloramphenicol
Kanamycin
Ampicillin
Bleocin(Zeocin)

Inventory

Well Plasmid Resistance
A / 1 pMXLC_pFa
Chloramphenicol
A / 2 pMXLC_aFb
Chloramphenicol
A / 3 pMXLC_bFc
Chloramphenicol
A / 4 pMXLC_cFd
Chloramphenicol
A / 5 pMXLC_dFe
Chloramphenicol
A / 6 pMXLC_aFq
Chloramphenicol
A / 7 pMXLC_bFq
Chloramphenicol
A / 8 pMXLC_cFq
Chloramphenicol
A / 9 pMXLC_dFq
Chloramphenicol
A / 10 pMXLC_eFq
Chloramphenicol
A / 11 pMXBC_pFa
Chloramphenicol
A / 12 pMXBC_aFb
Chloramphenicol
B / 1 pMXBC_bFc
Chloramphenicol
B / 2 pMXBC_aFq
Chloramphenicol
B / 3 pMXBC_bFq
Chloramphenicol
B / 4 pMXBC_cFq
Chloramphenicol
B / 5 pMXLK_pFa
Kanamycin
B / 6 pMXLK_aFb
Kanamycin
B / 7 pMXLK_bFc
Kanamycin
B / 8 pMXLK_cFd
Kanamycin
B / 9 pMXLK_dFe
Kanamycin
B / 10 pMXLK_aFq
Kanamycin
B / 11 pMXLK_bFq
Kanamycin
B / 12 pMXLK_cFq
Kanamycin
C / 1 pMXLK_dFq
Kanamycin
C / 2 pMXLK_eFq
Kanamycin
C / 3 pMXBK_pFa
Kanamycin
C / 4 pMXBK_aFb
Kanamycin
C / 5 pMXBK_bFc
Kanamycin
C / 6 pMXBK_aFq
Kanamycin
C / 7 pMXBK_bFq
Kanamycin
C / 8 pMXBK_cFq
Kanamycin
C / 9 pMXLP_pFa
Ampicillin
C / 10 pMXLP_aFb
Ampicillin
C / 11 pMXLP_bFc
Ampicillin
C / 12 pMXLP_cFd
Ampicillin
D / 1 pMXLP_dFe
Ampicillin
D / 2 pMXLP_aFq
Ampicillin
D / 3 pMXLP_bFq
Ampicillin
D / 4 pMXLP_cFq
Ampicillin
D / 5 pMXLP_dFq
Ampicillin
D / 6 pMXLP_eFq
Ampicillin
D / 7 pMXBP_pFa
Ampicillin
D / 8 pMXBP_aFb
Ampicillin
D / 9 pMXBP_bFc
Ampicillin
D / 10 pMXBP_aFq
Ampicillin
D / 11 pMXBP_bFq
Ampicillin
D / 12 pMXBP_cFq
Ampicillin
E / 1 DH10B-M.Osp807II
Bleocin(Zeocin)
E / 2 DH10B-M2.NmeMC58II
Bleocin(Zeocin)
E / 3 DH10B-M.XmnI
Bleocin(Zeocin)
Data calculated @ 2019-07-18

Kit Plate Map - #1000000153

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