STAR TALE Assembly Kit
(Kit #
1000000117
)
Depositing Lab: Steven Pollard
This kit offers users a TALE assembly reaction (STAR) method to construct multiple TALEs in a fast and simple manner. STAR enables production of TALENs, TALE-TFs and TALE-CEs. This kit contains a starting library of 68 fragment parts, allowing specific TALE repeat sequence generation within ~8 hours. Sequence-verified TALENs or TALE-TF plasmids targeting 17bp target sequences can be produced within three days, without the need for stepwise intermediate plasmid production. This kit also offers 16 pTALE-STAR destination vectors.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publications
STAR: a simple TAL effector assembly reaction using isothermal assembly. Gogolok S, Garcia-Diaz C, Pollard SM. Sci Rep. 2016 Sep 12;6:33209. doi: 10.1038/srep33209. PubMed PMID 27615025.
STAR: a simple TAL effector assembly reaction using isothermal assembly. Gogolok S, Kober U, Pollard SM. Methods in Mol Biol. 2018, 1772:477-490. doi: 10.1007/978-1-4939-7795-6_29. PubMed PMID 29754248.
Description
The STAR protocol allows quick and simple assembly of TAL effectors in three days based on single tube Gibson reactions. The kit is designed for assembly of DNA-binding domains of 16.5 RVDs and allows the use of any functional domain, ranging from nucleases to transcriptional activators and repressors.
Library generation is based on simple PCR reactions and no subcloning reactions of parts or destination vectors are needed.
Instead, 1mer pieces coding for single TALE repeat units, are assembled in a two-step Gibson reaction. Intermediate clean up steps are based on magnetic bead clean up protocols, which allows automation of the protocol if a higher throughput is required.
Summary of 8h assembly of TALE repeat domain with STAR protocol using two separate Gibson assembly reactions; 4×1mers, and 4×4mers plus vector. Please see Figure 1 from original publication for more details (Pubmed PMID 27615025).
Kit Documentation
STAR TALE Assembly Kit Protocol 28.8Â 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 STAR TALE Assembly Kit was a gift from Steven Pollard (Addgene kit #1000000117).”
For your Reference section:
STAR: a simple TAL effector assembly reaction using isothermal assembly. Gogolok S, Garcia-Diaz C, Pollard SM. Sci Rep. 2016 Sep 12;6:33209. doi: 10.1038/srep33209. PubMed PMID 27615025.
STAR: a simple TAL effector assembly reaction using isothermal assembly. Gogolok S, Kober U, Pollard SM. Methods in Mol Biol. 2018, 1772:477-490. doi: 10.1007/978-1-4939-7795-6_29. PubMed PMID 29754248.
STAR TALE Assembly Kit - #1000000117
- 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 |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | HD-1 | Ampicillin |
| A / 2 | HD-2 | Ampicillin |
| A / 3 | HD-3 | Ampicillin |
| A / 4 | HD-4 | Ampicillin |
| A / 5 | HD-5 | Ampicillin |
| A / 6 | HD-6 | Ampicillin |
| A / 7 | HD-7 | Ampicillin |
| A / 8 | HD-8 | Ampicillin |
| A / 9 | HD-9 | Ampicillin |
| A / 10 | HD-10 | Ampicillin |
| A / 11 | HD-11 | Ampicillin |
| A / 12 | HD-12 | Ampicillin |
| B / 1 | HD-13 | Ampicillin |
| B / 2 | HD-14 | Ampicillin |
| B / 3 | HD-15 | Ampicillin |
| B / 4 | HD-16 | Ampicillin |
| B / 5 | NI-1 | Ampicillin |
| B / 6 | NI-2 | Ampicillin |
| B / 7 | NI-3 | Ampicillin |
| B / 8 | NI-4 | Ampicillin |
| B / 9 | NI-5 | Ampicillin |
| B / 10 | NI-6 | Ampicillin |
| B / 11 | NI-7 | Ampicillin |
| B / 12 | NI-8 | Ampicillin |
| C / 1 | NI-9 | Ampicillin |
| C / 2 | NI-10 | Ampicillin |
| C / 3 | NI-11 | Ampicillin |
| C / 4 | NI-12 | Ampicillin |
| C / 5 | NI-13 | Ampicillin |
| C / 6 | NI-14 | Ampicillin |
| C / 7 | NI-15 | Ampicillin |
| C / 8 | NI-16 | Ampicillin |
| C / 9 | NG-1 | Ampicillin |
| C / 10 | NG-2 | Ampicillin |
| C / 11 | NG-3 | Ampicillin |
| C / 12 | NG-4 | Ampicillin |
| D / 1 | NG-5 | Ampicillin |
| D / 2 | NG-6 | Ampicillin |
| D / 3 | NG-7 | Ampicillin |
| D / 4 | NG-8 | Ampicillin |
| D / 5 | NG-9 | Ampicillin |
| D / 6 | NG-10 | Ampicillin |
| D / 7 | NG-11 | Ampicillin |
| D / 8 | NG-12 | Ampicillin |
| D / 9 | NG-13 | Ampicillin |
| D / 10 | NG-14 | Ampicillin |
| D / 11 | NG-15 | Ampicillin |
| D / 12 | NG-16 | Ampicillin |
| E / 1 | NH-1 | Ampicillin |
| E / 2 | NH-2 | Ampicillin |
| E / 3 | NH-3 | Ampicillin |
| E / 4 | NH-4 | Ampicillin |
| E / 5 | NH-5 | Ampicillin |
| E / 6 | NH-6 | Ampicillin |
| E / 7 | NH-7 | Ampicillin |
| E / 8 | NH-8 | Ampicillin |
| E / 9 | NH-9 | Ampicillin |
| E / 10 | NH-10 | Ampicillin |
| E / 11 | NH-11 | Ampicillin |
| E / 12 | NH-12 | Ampicillin |
| F / 1 | NH-13 | Ampicillin |
| F / 2 | NH-14 | Ampicillin |
| F / 3 | NH-15 | Ampicillin |
| F / 4 | NH-16 | Ampicillin |
| F / 5 | pTALEN-STAR-T | Ampicillin |
| F / 6 | pTALEN-STAR-A | Ampicillin |
| F / 7 | pTALEN-STAR-C | Ampicillin |
| F / 8 | pTALEN-STAR-G | Ampicillin |
| F / 9 | pTALE-STAR-T_RFP entry | Ampicillin |
| F / 10 | pTALE-STAR-A_RFP entry | Ampicillin |
| F / 11 | pTALE-STAR-C_RFP entry | Ampicillin |
| F / 12 | pTALE-STAR-G_RFP entry | Ampicillin |
| G / 1 | pTALE-STARVP64-T | Ampicillin |
| G / 2 | pTALE-STARVP64-A | Ampicillin |
| G / 3 | pTALE-STARVP64-C | Ampicillin |
| G / 4 | pTALE-STARVP64-G | Ampicillin |
| G / 5 | pTALE-STARVP160-T | Ampicillin |
| G / 6 | pTALE-STARVP160-A | Ampicillin |
| G / 7 | pTALE-STARVP160-C | Ampicillin |
| G / 8 | pTALE-STARVP160-G | Ampicillin |
| G / 9 | pTALE-STAR-KRAB-T | Ampicillin |
| G / 10 | pTALE-STAR-KRAB-A | Ampicillin |
| G / 11 | pTALE-STAR-KRAB-G | Ampicillin |
| G / 12 | pTALE-STAR-KRAB-C | Ampicillin |