(cDNA library) Human ; Umbilical Vein Endothelial Cells

code:02-721

This cDNA library (plasmid DNA) is constructed from Human Umbilical Vein Endothelial Cell (HUVEC)-derived poly(A)+ RNA by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express human genes in mammalian cells as it contains SV40 promoter. It also contains Ori of pUC plasmid required for replication in E.coli f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoters for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)

(cDNA library) Mouse ; Thymocyte

Code:02-713

This cDNA library (plasmid DNA) is constructed from mouse thymocyte-derived poly(A)+ RNA by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express genes in mammalian cells as it contains SV40 promoter. It also contains f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoter for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)

(cDNA library) Rat ; Rat embryonic fibroblast

code:02-717

This cDNA library (plasmid DNA) is constructed from rat embryonic fibroblast-derived poly(A)+ RNA by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express rat genes in mammalian cells as it contains SV40 promoter. It also contains Ori of pUC plasmid required for replication in E.coli f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoters for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)

Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 62 x 106
2) Average insert size : longer than 1 kb
Storage: -20℃

(cDNA library) Xenopus ; oocyte

code:02-711

This cDNA library (plasmid DNA) is constructed from Xenopus oocyte-derived poly(A)+ RNA by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pBA2 vector used in this library has pUC ori which enables replication in E. coli and Ampr as a selection marker.

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression level of mRNA of the particular gene.)

Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 1.1 x 106
2) Average insert size : longer than 1 kb
Storage: -20℃

cDNA Library S. cerevisiae Log Phase

code:02-701

This cDNA library (plasmid DNA) is constructed from Saccharomyces cerevisiae strain S288C- derived poly(A)+ RNA at the log phase by the Linker-Primer method　(Ref.1) by Prof. H. Nojima of Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pLZ3 vector (shown below) used in this library can not replicate in S. cereviseaas but contains pUCori for replication in E. coli

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)

Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 3.6 x 106
2) Average insert size : longer than 1 kb
Storage: -20℃

References
1. KoboriM. IkedaY. NaraH. KatoM. KumegawaM. NojimaH. and KawashimaH. ” Large scale isolation of osteoclast-specific genes by an improved method involving the preparation of a subtracted cDNA library.” Genes Cells 3: 459-475 (1998) PMID: 9753427
2. TanakaS. and NojimaH. ”Nik1: a Nim1-like protein kinase of S. cerevisiae interacts with the Cdc28 complex and regulates cell cycle progression.” Genes Cells 1 905-921 (1996) PMID: 9077450
3. SambrookJ. and RussellDW. Molecular Cloning Chapter 11 ”Preparation of cDNA libraries and gene identification.” CSHL Press (2001)

(cDNA library) S.pombe ; mitosis

code:02-703

This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe strain h-L972- derived poly(A)+ RNA at the log phase by the Linker-Primer method (Ref.1) by Prof. H. Nojima of Research Institute for Microbial Diseases Osaka University. cDNAs in this library are unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme sites of Not I and BamHI (Bgl II)-Sma I adaptor.
The pLZ3 vector used in this library can replicate both in S. pombe and E.coli and express the S. pombe genes in mammalian cells as it contains SV40 promoter as well as in S. pombe (see Figure and Ref.2).

Applications
1. PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector (Ref 3). The cloned cDNAs are useful for identifying the coding region large-scale protein production and preparation of probes etc. Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression levels of mRNA of the gene of interest.)
2. Cloning the cDNA by functional complementation of the corresponding S. pombe mutants.
Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 28 x 106
2) Average insert size: longer than 1 kb
Storage: -20℃

cDNA Library S.pombe; Meiosis

code:02-705

This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe strain CD16-1(h+/h-) derived poly(A)+ RNA at the state of miosis by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. cDNAs in this library are unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express the S. pombe genes in mammalian cells as it contains SV40 promoter. It also contains f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoter for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression levels of mRNA of the genes.)

Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 1.3 x 106
2) Average insert size : longer than 1 kb
Storage: -20℃

(cDNA library) Mouse ; Testis

code:02-715

This cDNA library (plasmid DNA) is constructed from mouse testis-derived poly(A)+ RNA by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express mouse genes in mammalian cells as it contains SV40 promoter. It also contains pUC plasmid Ori required for replication in E. coli and f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoter for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)

Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 6 x 106
2) Average insert size : longer than 1 kb
Storage: -20℃

cDNA Library Rat NRK Cell Log Phase

code:02-719

This cDNA library (plasmid DNA) is constructed from poly(A)+ RNA of rat NRK (normal rat kidney) cells at the 0hr log phase. It is constructed by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express rat genes in mammalian cells as it contains SV40 promoter. It also contains Ori of pUC plasmid required for replication in E.coli f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoters for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)
Specification
Quantity: 500 ng (40 ng/ul 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality: 1) Number of independent clones: 1.4 x 106
2) Average insert size : longer than 1 kb
Storage: -20℃

(cDNA library) Human ; HeLa (#1)

code:02-723

This cDNA library (plasmid DNA) is constructed from HeLa cell-derived poly(A)+ RNA by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. This library is unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor.
The pAP3neo vector used in this library can express human genes in mammalian cells as it contains SV40 promoter. It also contains Ori of pUC plasmid required for replication in E.coli f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoters for RNA synthesis (see Figure). GenBank Accession No. AB003468

Application
PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc.
Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression rate of mRNA of the objective gene.)