Escherichia coli
Protocols: co-immunoprecipitation of flag -tagged Hfq protein and its bound RNA species was harvested using magnetic beads conjugated with anti-flag antibodies. The beads were washed stringently and bound RNA was eluted and used for cDNA library preperation. UTI89 was grown overnight in LB medium at 37 degree C and used to infect a monolayer of bladder epithelial cell line PD07i. This was used as infection. RNA was eluted from anti-flag antibody conjegated magnetic beads by phenol-chloroform extraction.. The RNA samples were poly(A)-tailed using poly(A) polymerase, followed by treatment with tobacco acid pyrophosphatase (TAP). Then a RNA adapter was ligated to the 5 prime-phosphate of the RNA Firststrand cDNA synthesis was performed using an oligo(dT)-adapter primer and M-MLV reverse transcriptase. The resulting cDNAs were PCR-amplified to about 20-60 ng/uI using a high fidelity DNA polymerase. PCR cycles performed and barcode sequences, which are integrated within the 3 prime sequencing adaptor.
ENA first public
2016-05-31
ENA last update
2015-04-20
External Id
SAMEA3355461
INSDC center alias
Biochemistry and Molecular Biology Department, University of Southern Denmark, Odense, Denmark.
INSDC center name
Biochemistry and Molecular Biology Department, University of Southern Denmark, Odense, Denmark.
INSDC first public
2016-05-31T17:01:39Z
INSDC last update
2015-04-20T16:43:57Z
INSDC status
public
Submitter Id
E-MTAB-3498:Infection
broker name
ArrayExpress
sample name
E-MTAB-3498:Infection
strain
UTI89