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NASC

The European Arabidopsis Stock Centre

SAIL lines from Syngenta

Donated by

  • Alberto Martinez Jealott's Hill International Research Centre, Syngenta AG
  • Joseph R. Ecker Plant Biology Laboratory, The Salk Institute for Biological Studies
  • Allen Sessions Torrey Mesa Research Institute, Syngenta AG

Click here to view all 54699 of these lines.

Description

SAIL lines (Syngenta Arabidopsis Insertion Library)

The Syngenta Arabidopsis Insertion Library (SAIL) collection was generated from approximately 100,000 individual T-DNA mutagenised Arabidopsis plants (Columbia ecotype). A modified approach to Thermal Asymmetric Interlaced-Polymerase Chain Reaction (TAIL-PCR) was adopted to identify left border flanking sequences for these plants. The average number of T-DNA insertions per line is 1.5 to 2.0. The typical TAIL PCR border sequence from each plant is over 300 nucleotides in length and often contains sequence from two separate TAIL products.

The collection includes approximately 45,000 lines.

Constructs

Two different binary vectors were used to generate the SAIL population. The T-DNA in pCSA110 is 7541 kb in length and contains, from the left to right border, a BASTA resistance cassette, pBluescript SKII+, and a Lat52 promoter-B-glucuronidase fusion.

The T-DNA in pDAP101 is 4763 kb in length and contains, from the left to right border, a BASTA resistance cassette and a pBluescript SKII+. Both T-DNAs have the same left border but different right borders.

Confirmation of inserts

The left border (both T-DNA have the same left border sequence):

LB1 (5'-GCC TTT TCA GAA ATG GAT AAA TAG CCT TGC TTC C-3')
LB2 (5'-GCT TCC TAT TAT ATC TTC CCA AAT TAC CAA TAC A-3')
LB3 (5'-TAG CAT CTG AAT TTC ATA ACC AAT CTC GAT ACA C-3')

For the right border sequence in pCSA110:

qRB1 (5'-CAA ACT AGG ATA AAT TAT CGC GCG CGG TGT C-3')
qRB2 (5'-GGT GTC ATC TAT GTT ACT AGA TCG GGA ATT GA-3')
qRB3 (5'-CGC CAT GGC ATA TGC TAG CAT GCA TAA TTC-3')

For the right border sequence in pDAP101:

RB1 (5'-ATT AGG CAC CCC AGG CTT TAC ACT TTA TG-3')
RB2 (5'-GTA TGT TGT GTG GAA TTG TGA GCG GAT AAC-3')
RB3 (5'-TAA CAA TTT CAC ACA GGA AAC AGC TAT AC-3')
RB3 (5'-TAA CAA TTT CAC ACA GGA AAC AGC TAT AC-3')

References

  • McElver, J. et al. 2001. Insertional mutagenesis of genes required for seed development in Arabidopsis thaliana. Genetics 159(4):1751-63.PubMed ID: 11779812.
  • Sessions, A. et al. 2002. A high-throughput Arabidopsis reverse genetics system. The Plant cell 14(12):2985-94.PubMed ID: 12468722.