FLASH Assembly

FLASH is an automatable high-throughput method for assembling DNA encoding TAL effector repeat arrays developed by the Joung lab (Reyon & Tsai et al., Nat Biotechnol. 2012). With automated FLASH, DNA fragments encoding 96 variable-length TAL effector repeat arrays can be assembled in one day. When practiced manually with a multi-channel pipet, FLASH can also be used by a single researcher to make DNA fragments encoding 12 to 24 variable-length TAL effector repeat arrays in one to two days. TAL effector repeat arrays assembled by FLASH are identical in DNA sequence to those assembled by the REAL and REAL-Fast methods.

Protocols

For high-throughput (automated) FLASH assembly

For medium-throughput (manual) FLASH assembly

Important note: Although the published FLASH protocol describes assembly of DNA fragments on streptavidin-coated magnetic beads, the latest version of the method now performs assembly in streptavidin-coated wells of a 96-well plate. This updated protocol is available upon request from the Joung Lab.

Reagents (for both high- and medium-throughput FLASH)

Archive of 376 plasmids encoding pre-assembled TAL repeats (available by request from the Joung lab)

TALEN expression vectors from Addgene

Software (for both high- and medium-throughput FLASH)

ZiFiT Targeter
(publicly available, web-based program that identifies potential FLASH TALEN target sites and provides information about plasmids required to assemble specific TALEN using FLASH)

   

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