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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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