GSS clones
B. oleracea shotgun clones (TIGR)
Background
TIGR received funding, under Cooperative Agreement DBI-9813586 from the National Science Foundation, to shotgun sequence several whole genome libraries of Brassica oleracea genomic DNA .TIGR has produced approximately four hundred thousand (400,000) sequences and accumulated approximately two hundred thousand (200,000) small insert clones (referred to as "TIGR Brassica oleracea Clone Collection"). The sequences from these are avaliable from NCBI/GenBank.Availability
WarwickHRI now has the non-exclusive distribution rights of the TIGR Brassica oleracea Clone Collection. The sequenced shotgun B. oleracea clones from TIGR are now stored under archive conditions at HRI HRI is the sole supplier of these clones to the research community.Library stats
TIGR libary_id | DNA | Insert Size | Vector |
---|---|---|---|
BOG, BOH, BOM, BON | Whole plant | 2-3 kb | pHOS1 |
BOIA | Nuclear | 1.2-1.4 kb | pHOS2 |
BOIB-H | Nuclear | 1.4-1.6 kb | pHOS2 |
BOJ | Whole plant | 1.6-2 kb | pHOS2 |
BOK | Whole plant | 10-12 kb | pHOS2 |
The inserts are cloned into the BstX1 site and can be popped out with this enzyme. You should also be able to use combinations of EcoR1 + BamH1 or Pst1. It is not TIGR policy at the moment to release the entire sequence of their plasmids but the maps of the two used is available The maps of the plasmids used are attached. They are to all intents and purposes the same -just different origins of replication. Selection with these plasmids is using AMP.
Primers used and plasmid maps
The primers TIGR used for sequencing where:TIGR Universal Forward - 18 bp (TF) TGT AAA ACG ACG GCC AGT
TIGR Universal Reverse - 18 bp (TR) CAG GAA ACA GCT ATG ACC
N.B. (TIGR Universal Forward and reverse primers are M1f (-20) and M13r respectively)
The positions of these are shown on the plasmid maps pHOS1 and pHOS2