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New release: 19,000 additional SLAT stable transposon insertion lines.

Sean May [ NASC ] sean_may at thale.life.nottingham.ac.uk
Fri Oct 13 06:33:16 EST 2000

The fourth and fifth sets of lines from the SLAT stable transposon
insertion population are now available from http://arabidopsis.org.uk This
represents an additional 19,000 lines, (380 pools of 50 lines) bringing
the running total up to 47,250.


Seeds are available both as a set, and as separate pools of 50 lines.

Please note that all of the seeds stocks relating to the SINS database of
flanking insertion sequence tags are already available.
When ordering SINS hits, please order the 50 pool relating to the first
two numbers in the SINS database (e.g. a hit in 02_06_11 would be a hit in
pool 02_06 with individual sequence 11 - so 'search' Donor Stock No. 02_06
in the catalogue to order the 50 pool).

The SLAT lines were generated by Dr Jonathan Jones, Sainsbury Laboratory,
John Innes Centre, Norwich, UK

Seeds and DNA from these lines were bulked for distribution by Ottoline
Leyser, University of York, UK.

The SLAT population provides both a useful forward and reverse genetic
resource, especially as enhanced by the SINS database facility and by use
of SLAT filter hybridisation.

It is important that you should unambiguously confirm that the DNA from
the pool that you have ordered, tests positive for the insert location
that you expect to have received before expending time on the seed stocks.
See below under 'why order DNA'.

The background of this population is Columbia (Col-0 : Stock number
The binary vector used to generate the lines contains a T-DNA that carries
(i) a non-autonomous Spm derivative containing the BAR gene selectable
marker, (ii) a counter selectable genetic marker based on a bacterial
cytochrome P450 that activates a DuPont proherbicide and (iii) an Spm
transposase (TPase) gene under the control of the P35S, pSpm or
meiosis-specific pAtDMC1 promoters. This construct permits the selection
for stable, unlinked with T-DNA, single copy insertions.

Note: 1 set = 4 superpools; 1 superpool = 48 pools; 1 pool = 50 lines.
This year (Y2K) the released population will become 6 sets (~= 6 x 4 x 48
x 50 = 57,600 lines).

Further information about the Slat lines and links to the SINS database
are available from the Sainsbury laboratory Plant Molecular Genetics
webpage :
http://www.jic.bbsrc.ac.uk/sainsbury-lab/jonathan-jones/jjhome.htm and
many details about these lines can also be found also in our catalogue
pages at arabidopsis.org.uk under the appropriate 'browse' heading..


The seed pools are maintained and distributed as pools of 50 lines.

Set 4 constitutes 191 pools of 50 lines and can be ordered as stock number

Set 5 constitutes 189 pools of 50 lines and can be ordered as stock number

DNA for these sets will be released shortly.

Why order DNA ?

The superpools are useful as reverse genetics resources in order to
pinpoint potential positives, 4 PCRs + controls will screen 9,600
lines (4 x 48 x 50). The separate 50-pools are useful for confirming the
efficacy of PCR primers and probes designed prior to receiving the
materials, and reduces / eliminates the time necessary for growing
50-pools prior to confirming the hit. Also they allow the potential for
extending the sequencing of the insert flanking region for homologue
checking and sequence verification.


To ensure that the maximum number of individuals can obtain the lines as
easily as possible we have chosen to recover distribution costs in the
following way:
Each stock (DNA or seed) is taken to represent one normal NASC stock and
as such is subject to our 'occasional / low / high' user method of
charging (which we operate in parallel with the ABRC for other stocks).
This should encourage exploratory use of the lines and return our normal
level of cost recovery from more frequent users.

Each 'reverse genetic' superpool DNA stock has a charge of 15 UKP levied
against it.

Each set of 192 x 50 lines has a handling and distribution charge of 300
UKP associated with it.

Any queries concerning these lines or any other lines distributed by the
stock centres should be directed to arabidopsis at nasc.nott.ac.uk.

Sean_May at nasc.nott.ac.uk (D.Phil.)
Director, Nottingham Arabidopsis Stock Centre (NASC)
School of Biosciences, Plant Science Division,
University of Nottingham, Biological Sciences Building,
University Park, Nottingham. NG7 2RD. UK

FON:			+44 (0)115 9513237
FAX:			+44 (0)115 9513297
Catalogue:		seeds.arabidopsis.org.uk
Genomics:		ukcrop.net
Transcriptomics:	garnet.arabidopsis.org.uk
ICQ:			57288800


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