The FGSC holds the libraries used in the Neurospora genome program at the Broad Institute and can distribute individual clones or groups of clones according to genome location data (below). Full ordered libraries is not available without special arrangement.
These cosmids (and others) are mapped on assembly 12 of the Neurospora genome
Pricing and shipping information
The codes used on the Broad Website for EST clones refer to the libraries as follows:
G906- 7-hours Vegetative Growth
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Cells were grown in 1X Vogel’s medium/2% sucrose for 7 h at 34 degrees C with
orbital shaking at 125 rpm. First-strand cDNA containing methyl-C was
synthesized using an oligo-dT primer bearing a XhoI site to enable directional
cloning. After second-strand synthesis, addition of EcoRI adapters, digestion
with XhoI, and appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested
UNIZAP XR Lambda arms and the ligation products packaged into Lambda particles.
The Lambda cDNA library was amplified. Mass-excision of the amplified library
was accomplished to produce pBluescript phagemid for cDNA sequencing.
Baştürkmen-Sachs 02
G991- 24-hours Vegetative Growth
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Cells were grown in 1X Vogel’s medium/2% sucrose for 24 h at 30 degrees C with
orbital shaking at 200 rpm. First-strand cDNA containing methyl-C was
synthesized using an oligo-dT primer bearing a XhoI site to enable directional
cloning. After second-strand synthesis, addition of EcoRI adapters, digestion
with XhoI, and appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested
UNIZAP XR Lambda arms and the ligation products packaged into Lambda particles.
The Lambda cDNA library was amplified. Mass-excision of the amplified library
was accomplished to produce pBluescript phagemid for cDNA sequencing.
G992- 1-hour Heat Shock after 9-hours Vegetative Growth
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Cells were grown in 1X Vogel’s medium/2% sucrose for 9 h at 30 degrees C and
then for 1 h at 45 degrees C with orbital shaking at 200 rpm. First-strand cDNA
containing methyl-C was synthesized using an oligo-dT primer bearing a XhoI site
to enable directional cloning. After second-strand synthesis, addition of EcoRI
adapters, digestion with XhoI, and appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested
UNIZAP XR Lambda arms and the ligation products packaged into Lambda particles.
The Lambda cDNA library was amplified. Mass-excision of the amplified library
was accomplished to produce pBluescript phagemid for cDNA sequencing.
Baştürkmen-Sachs 04
G993- 1-hour Glucose Deprivation after 9-hours Vegetative Growth
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Cells were grown in 1X Vogel’s medium/2% glucose for 9 h at 30 degrees C with
orbital shaking at 200 rpm, and then for 1 h in 1X Vogel’s medium lacking
glucose. First-strand cDNA containing methyl-C was synthesized using an oligo-dT
primer bearing a XhoI site to enable directional cloning. After second-strand
synthesis, addition of EcoRI adapters, digestion with XhoI, and appropriate
work-up, cDNA was ligated to XhoI/EcoRI-digested UNIZAP XR Lambda arms and the
ligation products packaged into Lambda particles. The Lambda cDNA library was
amplified. Mass-excision of the amplified library was accomplished to produce
pBluescript phagemid for cDNA sequencing.
Baştürkmen-Sachs 05
G994- 1-hour Nitrogen Deprivation after 9 hours Vegetative Growth
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Cells were grown in 1X Vogel’s medium/2% sucrose for 9 h at 30 degrees C with
orbital shaking at 200 rpm, and then for 1 h in 1X Vogel’s medium lacking
ammonium nitrate. First-strand cDNA containing methyl-C was synthesized using an
oligo-dT primer bearing a XhoI site to enable directional cloning. After
second-strand synthesis, addition of EcoRI adapters, digestion with XhoI, and
appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested UNIZAP XR Lambda
arms and the ligation products packaged into Lambda particles. The Lambda cDNA
library was amplified. Mass-excision of the amplified library was accomplished
to produce pBluescript phagemid for cDNA sequencing.
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Cells were grown in 1X Vogel’s medium/2% sucrose for 9 h at 30 degrees C with
orbital shaking at 200 rpm; sodium chloride was added to a final concentration
of 0.68M and incubation continued for 1 h. First-strand cDNA containing methyl-C
was synthesized using an oligo-dT primer bearing a XhoI site to enable
directional cloning. After second-strand synthesis, addition of EcoRI adapters,
digestion with XhoI, and appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested
UNIZAP XR Lambda arms and the ligation products packaged into Lambda particles.
The Lambda cDNA library was amplified. Mass-excision of the amplified library
was accomplished to produce pBluescript phagemid for cDNA sequencing.
G1176- 1-hour Oxidative Stress after 9-hours Vegetative Growth
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225).
Four separate cultures were incubated in 1X Vogel’s/1% sorbose/0.1% sucrose for
9 h at 30 degrees C with orbital shaking at 200 rpm, and then to each was added
(final concentrations indicated) either (1) 10 mM hydrogen peroxide (2) 750
micromolar sodium arsenite (3) 2.0 mM dithiothreitol and (4) 50 micromolar
cadmium chloride and incubation was continued for 1 h. First-strand cDNA
containing methyl-C was synthesized using an oligo-dT primer bearing a XhoI site
to enable directional cloning. After second-strand synthesis, addition of EcoRI
adapters, digestion with XhoI, and appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested
UNIZAP XR Lambda arms and the ligation products packaged into Lambda particles.
The Lambda cDNA library was amplified. After the mass-excision of the amplified
libraries equal titers of the four phagemid libraries were combined to produce
pBluescript phagemid for cDNA sequencing.
Baştürkmen-Sachs 11/12
G1175- 48-hours Unfertilized Growth in Crossing Medium
Poly(A) mRNA was purified from the Mauriceville-1c mat A strain (FGSC 2225). Two
cultures were grown in Westergaard’s medium (synthetic cross medium) for 48 h at
25 degrees C, one with orbital shaking at 200 rpm, and one without shaking.
First-strand cDNA containing methyl-C was synthesized from each using an
oligo-dT primer bearing a XhoI site to enable directional cloning. After
second-strand synthesis, addition of EcoRI adapters, digestion with XhoI, and
appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested UNIZAP XR Lambda
arms and the ligation products packaged into Lambda particles. The Lambda cDNA
library was amplified. After the mass-excision of the amplified libraries equal
titers of two phagemid libraries were combined to produce pBluescript phagemid
for cDNA sequencing.
Poly(A) mRNA was purified from a 7 day crossing-culture of Mauriceville-1c mat A
(FGSC 2225)crossed with ORS mat a (FGSC 2490). Cells were grown in Westergaard's
medium for 5 days prior to initiating crossing. First-strand cDNA containing
methyl-C was synthesized using an oligo-dT primer bearing a XhoI site to enable
directional cloning. After second-strand synthesis, addition of EcoRI adapters,
digestion with XhoI, and appropriate work-up, cDNA was ligated to XhoI/EcoRI-digested
UNIZAP XR Lambda arms and the ligation products packaged into Lambda particles.
The Lambda cDNA library was amplified. Mass-excision of the amplified library
was accomplished to produce pBluescript phagemid for cDNA sequencing.
Genomic libraries
cDNA libraries
Two cDNA libraries, which represent mRNA from the Neurospora crassa wild type strain, 74-OR23-1VA, were constructed in Lambda Zap version I+
The germinating conidia library is no longer available.