Platinum Gate TALEN Kit
(Kit #
1000000043
)
Depositing Lab: Takashi Yamamoto
This kit enables construction of highly-active Platinum TALENs using two-step Golden Gate cloning method. Platinum TALENs have variable TALE repeats with one of two types of TALE scaffold: +136/+63 or +153/+47. The kit contains 35 plasmdids: 16 module plasmids with non-RVD variations, 11 intermediate arrays, and 8 destination vectors.
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 from each plasmid page and will be shipped as bacterial stabs.
Original Publication
Repeating pattern of non-RVD variations in DNA-binding modules enhances TALEN activity. Sakuma T, Ochiai H, Kaneko T, Mashimo T, Tokumasu D, Sakane Y, Suzuki K, Miyamoto T, Sakamoto N, Matsuura S, Yamamoto T. Sci Rep 2013 Nov 29;3:3379. doi: 10.1038/srep03379. Pubmed PMID 24287550.
Description
The Platinum Gate TALEN kit enables construction of highly-active Platinum TALENs using two-step Golden Gate cloning method. Platinum TALENs have variable TALE repeats with one of two types of TALE scaffold: +136/+63 or +153/+47. The kit contains 16 module plasmids with non-RVD variations, 11 intermediate arrays, and 8 destination vectors.
These Platinum TALENs have been proven to work well in:
- cultured cells (doi: 10.1038/srep03379 and doi: 10.1073/pnas.1317008111)
- nematodes (doi: 10.1111/dgd.12108)
- sea urchins (doi: 10.1111/dgd.12099)
- ascidians (doi: 10.1242/dev.099572)
- newts (doi: 10.1111/dgd.12103)
- frogs (doi: 10.1111/dgd.12105 and doi: 10.1038/srep03379)
- mice (doi: 10.1538/expanim.63.79)
- rats (doi: 10.1038/srep03379)
Overview of TALEN assembly
Figure from Sakuma et al., Scientific Reports 2013 Nov 29;3:3379
Protocol
Download a PDF Protocol for TALEN construction using the Platinum Gate TALEN Kit as described in the original publication.
Platinum Gate TALEN assembly protocol 379.6Â KBResources
Download a ZIP file containing all Platinum Gate TALEN plasmid sequences in GenBank format
Platinum Gate TALEN Kit plasmids - GenBank files 128.0Â KBHow 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 TALEN kit used for constructing Platinum TALENs was a gift from Takashi Yamamoto (Addgene kit # 1000000043)"
For your Reference section:
Repeating pattern of non-RVD variations in DNA-binding modules enhances TALEN activity. Sakuma T, Ochiai H, Kaneko T, Mashimo T, Tokumasu D, Sakane Y, Suzuki K, Miyamoto T, Sakamoto N, Matsuura S, Yamamoto T. Sci Rep 2013 Nov 29;3:3379. doi: 10.1038/srep03379. PubMed
PMID 24287550.
Platinum Gate TALEN Kit - #1000000043
- 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
| Ampicillin | |
| Spectinomycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | p1HD | Ampicillin |
| A / 2 | p1NG | Ampicillin |
| A / 3 | p1NI | Ampicillin |
| A / 4 | p1NN | Ampicillin |
| A / 5 | p2HD | Ampicillin |
| A / 6 | p2NG | Ampicillin |
| A / 7 | p2NI | Ampicillin |
| A / 8 | p2NN | Ampicillin |
| A / 9 | p3HD | Ampicillin |
| A / 10 | p3NG | Ampicillin |
| A / 11 | p3NI | Ampicillin |
| A / 12 | p3NN | Ampicillin |
| B / 1 | p4HD | Ampicillin |
| B / 2 | p4NG | Ampicillin |
| B / 3 | p4NI | Ampicillin |
| B / 4 | p4NN | Ampicillin |
| B / 5 | ptCMV-136/63-VR-HD | Ampicillin |
| B / 6 | ptCMV-136/63-VR-NG | Ampicillin |
| B / 7 | ptCMV-136/63-VR-NI | Ampicillin |
| B / 8 | ptCMV-136/63-VR-NN | Ampicillin |
| B / 9 | ptCMV-153/47-VR-HD | Ampicillin |
| B / 10 | ptCMV-153/47-VR-NG | Ampicillin |
| B / 11 | ptCMV-153/47-VR-NI | Ampicillin |
| B / 12 | ptCMV-153/47-VR-NN | Ampicillin |
| C / 1 | pFUS2_a1a | Spectinomycin |
| C / 2 | pFUS2_a2a | Spectinomycin |
| C / 3 | pFUS2_a2b | Spectinomycin |
| C / 4 | pFUS2_a3a | Spectinomycin |
| C / 5 | pFUS2_a3b | Spectinomycin |
| C / 6 | pFUS2_a4a | Spectinomycin |
| C / 7 | pFUS2_a4b | Spectinomycin |
| C / 8 | pFUS2_b1 | Spectinomycin |
| C / 9 | pFUS2_b2 | Spectinomycin |
| C / 10 | pFUS2_b3 | Spectinomycin |
| C / 11 | pFUS2_b4 | Spectinomycin |