New release: 9,050 additional SLAT stable transposon insertion lines.

Sean May [NASC] sean_may at thale.life.nottingham.ac.uk
Thu Jan 6 15:09:09 EST 2000


The third set of lines from the SLAT stable transposon insertion population
are now available from http://nasc.nott.ac.uk. or
http://seeds.nottingham.ac.uk
This represents an additional 9,050 lines (181 pools of 50 lines) bringing
the running total up to 28,250.
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Seeds are available both as a set, and as separate pools of 50 lines.
Out of the theoretical 192 pools of set 3, a total of 11 of the 50 pools are
not available.

DNA for the separate pools of 50 and superpools of set 3 are *not currently
available* but are expected to be released in the near future.
DNA for the separate pools of 50 and for the superpools (4 superpools per
set) are available from the catalogue for both of the earlier sets 1 and 2.

Set 3 (4 superpools designated 09 to 11 plus 13), together with the first 2
sets (8 superpools designated 01 to 08), cover the currently required pools
for all requests from the SINS database of flanking insertion sequence tags.

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).
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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

The background of this population is Columbia (Col-0 : Stock number N1092).
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.
(During Y2K, the released population will become 5 sets = 5 x 4 x 48 x 50 =
48,000 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
-------------------------------------------------
Seed
The seed pools are maintained and distributed as pools of 50 lines. The set
3 release constitutes 181 pools of 50 lines and can be ordered with the
Stock Number N41982.

DNA for set 3 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.

Pricing.
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
Director, Nottingham Arabidopsis Stock Centre (NASC)
University Park, Nottingham University. U.K. NG7 2RD.
-------------------------------------------------------
FON:                    +44 (0)115 9513237
FAX:                    +44 (0)115 9513297
Catalogue:              nasc.nott.ac.uk
Genome Databases:       synteny.nott.ac.uk


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