GABI-Kat
Donated by
- Bernd Weisshaar CeBiTec (Center for Biotechnology), Faculty of Biology, Bielefeld University
Description
GABI-Kat: generation of Flanking Sequence Tags (FSTs) from T-DNA mutagenised A. thaliana plants (accession Columbia)
PLEASE ORDER A GABI-KAT SET AND *NOT* THE INDIVIDUAL STOCKS. ALL AVAILABLE INDIVIDUAL STOCKS ARE INCLUDED IN EACH SET AND YOU WILL NEED ALL OF THEM TO DETERMINE WHICH ONES ARE HETEROZYGOUS, HOMOZYGOUS OR WILD-TYPE.
GABI-KAT HAVE CONFIRMED THAT WITHIN EACH SET THERE IS STATISTICALLY AT LEAST ONE HOMOZYGOUS STOCK BUT THEY DO NOT KNOW *WHICH* INDIVIDUAL IT IS.
THEREFORE YOU NEED TO DETERMINE THIS YOURSELF.
ALL SETS BEGIN WITH N4. INDIVIDUAL LINES BEGIN WITH EITHER N2, N3 or N7.
当你订购一个GABI－KAT 系列，请选择一整个系列而不是个别样本. 这是因为如果你想确认哪一个是杂合的，纯合的还是野生型的，你必须拥有所有的独立样本. GABI-KAT已经确认每一个系列里面都会有一个是纯合的，但是他们不知道是哪一个，所以你必须自己去选择. 所有的系列都是以N4开始的，而独立样本是以N2,N3或N7开始的.
In other words: each NASC stock within a GABI-Kat set is an individual T3 seed stock derived from a single T2 plant, and each T2 plant that gave rise to an individual stock within a GABI-Kat set might have been heterozygous or homozygous or wild type for the insertion. Based on this, we recommend our customers to order complete sets and check whether the stocks they receive are heterozygous, homozygous or wild type. The donation of segregating families for each line allows direct access to potentially homozygous material for a given insertion line.
GABI-Kat was initiated at the Max-Planck-Institute of Plant Breeding Research and aims to generate at least 80000 T-DNA transformants (sequenced transformant available at NASC as a segregating set) for the accession Columbia (Col-0). Seeds of single lines (T2 seeds from T1 plants) were harvested, and genomic DNA from T1 plants was extracted to produce FSTs from their insertion sites by a linker-ligation-PCR based approach. The FST data were then mapped to the genome and the results have been made available both via the internet at the GABI-Kat site, and as a bulk download.
GABI-Kat lines available to order at NASC
The GABI-Kat lines are now available via NASC at standard user fees (standard fee per line and £15 per set) to the international research community. GABI-Kat lines that have been confirmed will be transferred to NASC as sets of T3 seed harvested from a segregating family of single T2 plants. It is recommended that the set is ordered as each line within a set may be segregating for the insert. This will allow direct user access to potentially homozygous material for a given insertion line.
Explanation of line numbering system
Each set of T3 seeds has been produced from their respective GK-****** parental line. This parental line number is also the name given to the set. Each set and individual line also has its own NASC number. For example, the table below shows the set GK-067H03 and the individual lines that make up this set.
If you order a set you will receive all individual lines that make up the set. So in the example below if you order the set you will receive 12 individual lines for the price of £15.
| Name | NASC code | Set/Individual line | Price £ |
|---|---|---|---|
| GK-067H03 | N406423 | Set | 15.00 |
| GK-067H03.01 | N300001 | Ind. line | 7.50 |
| GK-067H03.02 | N300002 | Ind. line | 7.50 |
| GK-067H03.03 | N300003 | Ind. line | 7.50 |
| GK-067H03.04 | N300004 | Ind. line | 7.50 |
| GK-067H03.05 | N300005 | Ind. line | 7.50 |
| GK-067H03.06 | N300006 | Ind. line | 7.50 |
| GK-067H03.07 | N300007 | Ind. line | 7.50 |
| GK-067H03.08 | N300008 | Ind. line | 7.50 |
| GK-067H03.09 | N300009 | Ind. line | 7.50 |
| GK-067H03.10 | N300010 | Ind. line | 7.50 |
| GK-067H03.11 | N300011 | Ind. line | 7.50 |
| GK-067H03.12 | N300012 | Ind. line | 7.50 |
Segregation counts
Three numbers are counted from the selection plates:
For example - 100:92:75
total: total number of seeds on the plate.
germinated: number of seeds that show signs of a root tip.
resistant: seedlings that grow on the selection plate.
Selection was done using 5.25 mg/L sulfadiazine where wild type plants bleached and did not generate secondary leaves.
Order
Search the NASC catalogue for the GABI-Kat name e.g. "GK-067H03" (including the quotation marks). This will display the set and individual lines produced from the parental line.