Structured Review

TaKaRa complementary dna cdna
Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from <t>cDNA</t> synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes <t>DNA</t> marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).
Complementary Dna Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 283 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Virus-Like Nanoparticle Vaccine Confers Protection against Toxoplasma gondii"

Article Title: Virus-Like Nanoparticle Vaccine Confers Protection against Toxoplasma gondii

Journal: PLoS ONE

doi: 10.1371/journal.pone.0161231

Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from cDNA synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes DNA marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).
Figure Legend Snippet: Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from cDNA synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes DNA marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).

Techniques Used: Polymerase Chain Reaction, Amplification, Synthesized, Marker, Clone Assay, Plasmid Preparation

2) Product Images from "Implication of the env Gene of the Human Endogenous Retrovirus W Family in the Expression of BDNF and DRD3 and Development of Recent-Onset Schizophrenia"

Article Title: Implication of the env Gene of the Human Endogenous Retrovirus W Family in the Expression of BDNF and DRD3 and Development of Recent-Onset Schizophrenia

Journal: Schizophrenia Bulletin

doi: 10.1093/schbul/sbp166

Human Endogenous Retrovirus (HERV)-W env Upregulates the Expression of Messenger RNA (mRNA) of Brain-Derived Neurotrophic Factor (BDNF), NTRK2, and DRD3 in Human U251 Glioma Cells. (A) BDNF, NTRK2, and DRD3 gene expression in the human U251 glioma cells. A 363-bp Complementary DNA fragment for BDNF mRNA, a 384-bp fragment for NTRK2 mRNA. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control. (B) Histograms show BDNF, NTRK2, and DRD3 mRNA levels quantified; each bar represents the mean ± standard error of the mean of 3 values of IL-8 amplification products normalized to the starting total RNA volumes and referred to the corresponding GADPH values. n = 3 (* P
Figure Legend Snippet: Human Endogenous Retrovirus (HERV)-W env Upregulates the Expression of Messenger RNA (mRNA) of Brain-Derived Neurotrophic Factor (BDNF), NTRK2, and DRD3 in Human U251 Glioma Cells. (A) BDNF, NTRK2, and DRD3 gene expression in the human U251 glioma cells. A 363-bp Complementary DNA fragment for BDNF mRNA, a 384-bp fragment for NTRK2 mRNA. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control. (B) Histograms show BDNF, NTRK2, and DRD3 mRNA levels quantified; each bar represents the mean ± standard error of the mean of 3 values of IL-8 amplification products normalized to the starting total RNA volumes and referred to the corresponding GADPH values. n = 3 (* P

Techniques Used: Expressing, Derivative Assay, Amplification

3) Product Images from "Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification"

Article Title: Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification

Journal: Journal of Virological Methods

doi: 10.1016/j.jviromet.2008.01.023

Electrophoretic analysis of cross-reaction in the PCV2 LAMP assay. PCV2-BJ, PCV1, PPV, PRV, and PRRSV were used as targets for the PCV2 LAMP assay. From left to right: lane M, DNA Marker DL-2000; lane 1, DNA of PCV2-BJ; lane 2, DNA of PCV1; lane 3, DNA of PPV; lane 4, DNA of PRV; lane 5, cDNA of PRRSV; lane 6, DNA from healthy swine tissue.
Figure Legend Snippet: Electrophoretic analysis of cross-reaction in the PCV2 LAMP assay. PCV2-BJ, PCV1, PPV, PRV, and PRRSV were used as targets for the PCV2 LAMP assay. From left to right: lane M, DNA Marker DL-2000; lane 1, DNA of PCV2-BJ; lane 2, DNA of PCV1; lane 3, DNA of PPV; lane 4, DNA of PRV; lane 5, cDNA of PRRSV; lane 6, DNA from healthy swine tissue.

Techniques Used: Lamp Assay, Marker

4) Product Images from "Opsin gene duplication and diversification in the guppy, a model for sexual selection"

Article Title: Opsin gene duplication and diversification in the guppy, a model for sexual selection

Journal: Proceedings of the Royal Society B: Biological Sciences

doi: 10.1098/rspb.2006.3707

Genomic DNA blots. ( a ) Genomic DNA from females (F) and males (M) of the Wild Istanbul strain digested with the indicated enzymes and hybridized with probes for opsins RH1, RH2-1, LWS and SWS1. A BamHI site in the RH2-1 cDNA sequence results in two RH2-1 opsin bands. ( b ) Pooled DNA of guppies from the Tranquille (Tr), Cumaná (C) and Quare (Qu) strains digested with HindIII and hybridized with a LWS probe. ( c ) DNA from individual females and males of indicated strains digested with PvuII and hybridized with a LWS probe.
Figure Legend Snippet: Genomic DNA blots. ( a ) Genomic DNA from females (F) and males (M) of the Wild Istanbul strain digested with the indicated enzymes and hybridized with probes for opsins RH1, RH2-1, LWS and SWS1. A BamHI site in the RH2-1 cDNA sequence results in two RH2-1 opsin bands. ( b ) Pooled DNA of guppies from the Tranquille (Tr), Cumaná (C) and Quare (Qu) strains digested with HindIII and hybridized with a LWS probe. ( c ) DNA from individual females and males of indicated strains digested with PvuII and hybridized with a LWS probe.

Techniques Used: Sequencing

5) Product Images from "The Mitochondrial Endonuclease M20 Participates in the Down-Regulation of Mitochondrial DNA in Pollen Cells"

Article Title: The Mitochondrial Endonuclease M20 Participates in the Down-Regulation of Mitochondrial DNA in Pollen Cells

Journal: Plant Physiology

doi: 10.1104/pp.18.00754

Generation of the atm20 mutant using CRISPR/Cas9 technology. A, Schematic diagram of the CRISPR construct containing a Cas9 expression cassette driven by an enhanced CaMV 35S promoter and an sgRNA controlled by the AtU6-26 promoter. B, A 4-bp deletion was detected in genomic DNA and cDNA of a T1 transformant (d4 mutation). The target sequence is shaded in blue, and the (PCI/PINT associated module) PAM domain is shaded in red. C, The 4-bp deletion causes a frame-shift within AtM20 and may lead to early translational termination. D, A smaller product (AtM20Δ) of AtM20 due to base deletion and early termination was detected in plants with the d4 mutation (top), and the mutation inactivates its nucleolytic activity in vitro (bottom). Arrowheads indicate the molecular products of AtM20 and AtM20Δ.
Figure Legend Snippet: Generation of the atm20 mutant using CRISPR/Cas9 technology. A, Schematic diagram of the CRISPR construct containing a Cas9 expression cassette driven by an enhanced CaMV 35S promoter and an sgRNA controlled by the AtU6-26 promoter. B, A 4-bp deletion was detected in genomic DNA and cDNA of a T1 transformant (d4 mutation). The target sequence is shaded in blue, and the (PCI/PINT associated module) PAM domain is shaded in red. C, The 4-bp deletion causes a frame-shift within AtM20 and may lead to early translational termination. D, A smaller product (AtM20Δ) of AtM20 due to base deletion and early termination was detected in plants with the d4 mutation (top), and the mutation inactivates its nucleolytic activity in vitro (bottom). Arrowheads indicate the molecular products of AtM20 and AtM20Δ.

Techniques Used: Mutagenesis, CRISPR, Construct, Expressing, Sequencing, Activity Assay, In Vitro

6) Product Images from "Ultrasensitive Detection of RNA and DNA Viruses Simultaneously Using Duplex UNDP-PCR Assay"

Article Title: Ultrasensitive Detection of RNA and DNA Viruses Simultaneously Using Duplex UNDP-PCR Assay

Journal: PLoS ONE

doi: 10.1371/journal.pone.0141545

Comparison of different probes-functionalized MMPs and oligos-functionalized AuNPs. (A) Functional magnetic microparticles MMP-p1, -p2, -p3, -p4, -p5 and -p6 were incubated with RNA of TGEV in hybridization buffer at 40°C for 30 minutes, followed by washing and magnetic separation. Then the MMP-RNA complexes were reverse transcribed into cDNA, which were detected by TGEV-specific RT-PCR. M: Trans 2K Plus DNA Marker; 1: MMP1; 2: MMP2; 3: MMP3; 4: MMP4; 5: MMP5; 6: MMP6. (B) TGEV RNA was incubated with oligo1, oligo2, oligo3, oligo4, oligo5 or oligo6 functionalized Au-NPs at 50°C for 40 min. Then the complexes were washed and precipitated by centrifugation, followed by reverse transcription and TGEV specific RT-PCR detection. M: Trans 2K Plus DNA Marker; 1: oligo1; 2: oligo2; 3: oligo3; 4: oligo4; 5: oligo5; 6: oligo6.
Figure Legend Snippet: Comparison of different probes-functionalized MMPs and oligos-functionalized AuNPs. (A) Functional magnetic microparticles MMP-p1, -p2, -p3, -p4, -p5 and -p6 were incubated with RNA of TGEV in hybridization buffer at 40°C for 30 minutes, followed by washing and magnetic separation. Then the MMP-RNA complexes were reverse transcribed into cDNA, which were detected by TGEV-specific RT-PCR. M: Trans 2K Plus DNA Marker; 1: MMP1; 2: MMP2; 3: MMP3; 4: MMP4; 5: MMP5; 6: MMP6. (B) TGEV RNA was incubated with oligo1, oligo2, oligo3, oligo4, oligo5 or oligo6 functionalized Au-NPs at 50°C for 40 min. Then the complexes were washed and precipitated by centrifugation, followed by reverse transcription and TGEV specific RT-PCR detection. M: Trans 2K Plus DNA Marker; 1: oligo1; 2: oligo2; 3: oligo3; 4: oligo4; 5: oligo5; 6: oligo6.

Techniques Used: Functional Assay, Incubation, Hybridization, Reverse Transcription Polymerase Chain Reaction, Marker, Centrifugation

7) Product Images from "Cloning and characterization of TaVIP2 gene from Triticum aestivum and functional analysis in Nicotiana tabacum"

Article Title: Cloning and characterization of TaVIP2 gene from Triticum aestivum and functional analysis in Nicotiana tabacum

Journal: Scientific Reports

doi: 10.1038/srep37602

Copy number and chromosomal location analysis of TaVIP2 in wheat genome by Southern blot. Three copies of TaVIP2 genes were present in the genome of common wheat revealed by various enzymes ( a ); genomic DNA of wheat line Luivo was digested by Eco RV, Sac I, Kpn I, and Dra I (lanes 1–4); genomic DNA of wheat line CB037 was digested by Eco RV, Sac I, Kpn I, and Dra I (lanes 5–8); plasmid DNA containing TaVIP2 was used as check (lane 9); there is no restriction site for Eco RV, Sac I, Kpn I, or Dra I in the genomic DNA sequence of TaVIP2 ; the full length cDNA of TaVIP2 labeled with Digoxigenin was used as the probe. One TaVIP2 gene was proved to be located on chromosome 1 A using seven durum wheat substitution lines analyzed by Southern blot ( b ); durum wheat substitution lines 1D(1 A), 2D(2 A), 3D(3 A), 4D(4 A), 5D(5 A), 6D(6 A), and 7D(7 A) in which a pair of A-genome chromosomes in durum wheat were replaced by its corresponding pair of D-genome chromosome from hexaploid wheat cultivar CS (lanes 1–7); receptor durum wheat line of Langdon was used as control (lane 8); plant DNA samples were digested with Dra I and then hybridized with the TaVIP2 coding sequence labeled with Digoxigenin as the probe; Substitution line 1D(1 A) showed a different band from the parent durum wheat line Langdon and the other six durum wheat substitution lines.
Figure Legend Snippet: Copy number and chromosomal location analysis of TaVIP2 in wheat genome by Southern blot. Three copies of TaVIP2 genes were present in the genome of common wheat revealed by various enzymes ( a ); genomic DNA of wheat line Luivo was digested by Eco RV, Sac I, Kpn I, and Dra I (lanes 1–4); genomic DNA of wheat line CB037 was digested by Eco RV, Sac I, Kpn I, and Dra I (lanes 5–8); plasmid DNA containing TaVIP2 was used as check (lane 9); there is no restriction site for Eco RV, Sac I, Kpn I, or Dra I in the genomic DNA sequence of TaVIP2 ; the full length cDNA of TaVIP2 labeled with Digoxigenin was used as the probe. One TaVIP2 gene was proved to be located on chromosome 1 A using seven durum wheat substitution lines analyzed by Southern blot ( b ); durum wheat substitution lines 1D(1 A), 2D(2 A), 3D(3 A), 4D(4 A), 5D(5 A), 6D(6 A), and 7D(7 A) in which a pair of A-genome chromosomes in durum wheat were replaced by its corresponding pair of D-genome chromosome from hexaploid wheat cultivar CS (lanes 1–7); receptor durum wheat line of Langdon was used as control (lane 8); plant DNA samples were digested with Dra I and then hybridized with the TaVIP2 coding sequence labeled with Digoxigenin as the probe; Substitution line 1D(1 A) showed a different band from the parent durum wheat line Langdon and the other six durum wheat substitution lines.

Techniques Used: Southern Blot, Plasmid Preparation, Sequencing, Labeling

8) Product Images from "Photosystem II antenna complexes CP26 and CP29 are essential for nonphotochemical quenching in Chlamydomonas reinhardtii, et al. Photosystem II antenna complexes CP26 and CP29 are essential for nonphotochemical quenching in Chlamydomonas reinhardtii"

Article Title: Photosystem II antenna complexes CP26 and CP29 are essential for nonphotochemical quenching in Chlamydomonas reinhardtii, et al. Photosystem II antenna complexes CP26 and CP29 are essential for nonphotochemical quenching in Chlamydomonas reinhardtii

Journal: Plant, Cell & Environment

doi: 10.1111/pce.13680

CP26‐ and CP29‐targeted gene disruption in Chlamydomonas reinhardtii by CRISPR/Cas9 genome editing. (a) DNA sequence alignment of wild type (Wt) and mutants on cp26 ( k6 ) and cp29 ( k9 ) obtained by CRISPR/Cas9 genome editing. The 20‐bp target sequence of sgRNA is reported before the red‐coloured PAM sequence. In the case of k9 mutant, insertions ( k9‐1 ) or deletions were induced ( k9‐2 ) by non‐homologous repair, whereas in the case of k6 mutant, a Cas9‐mediated hygromycin resistance cassette insertion was obtained in the target site. Double‐mutant k69 was obtained by hygromycin resistance cassette insertion in a k9 background. (b) PCR amplification of cp26 and cp29 CDS sequences from Wt, k6 , k9 , and k69 cDNA. In the case of k6 and k69 mutants, the cp26 CDS cannot be amplified due to hygromycin cassette insertion. (c) SDS‐PAGE analysis of Wt and mutant strains performed with the Tris‐Tricine buffer system. Fifteen microgram of Chl was loaded in each lane. Molecular weight (MW) markers are indicated on the left. CP26 and CP29 bands are marked. (d) Immunoblot with specific antibodies directed against CP26 and CP29 on the same lanes of (c). Immunoblot against CP43 was added as control of the loading. (e) qRT‐PCR on cp26 and cp29 genes in Wt and mutant strains. rack1 gene was used as control for qRT‐PCR (see details in Figure S3 )
Figure Legend Snippet: CP26‐ and CP29‐targeted gene disruption in Chlamydomonas reinhardtii by CRISPR/Cas9 genome editing. (a) DNA sequence alignment of wild type (Wt) and mutants on cp26 ( k6 ) and cp29 ( k9 ) obtained by CRISPR/Cas9 genome editing. The 20‐bp target sequence of sgRNA is reported before the red‐coloured PAM sequence. In the case of k9 mutant, insertions ( k9‐1 ) or deletions were induced ( k9‐2 ) by non‐homologous repair, whereas in the case of k6 mutant, a Cas9‐mediated hygromycin resistance cassette insertion was obtained in the target site. Double‐mutant k69 was obtained by hygromycin resistance cassette insertion in a k9 background. (b) PCR amplification of cp26 and cp29 CDS sequences from Wt, k6 , k9 , and k69 cDNA. In the case of k6 and k69 mutants, the cp26 CDS cannot be amplified due to hygromycin cassette insertion. (c) SDS‐PAGE analysis of Wt and mutant strains performed with the Tris‐Tricine buffer system. Fifteen microgram of Chl was loaded in each lane. Molecular weight (MW) markers are indicated on the left. CP26 and CP29 bands are marked. (d) Immunoblot with specific antibodies directed against CP26 and CP29 on the same lanes of (c). Immunoblot against CP43 was added as control of the loading. (e) qRT‐PCR on cp26 and cp29 genes in Wt and mutant strains. rack1 gene was used as control for qRT‐PCR (see details in Figure S3 )

Techniques Used: CRISPR, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification, SDS Page, Molecular Weight, Quantitative RT-PCR

9) Product Images from "The Mitochondrial Endonuclease M20 Participates in the Down-Regulation of Mitochondrial DNA in Pollen Cells"

Article Title: The Mitochondrial Endonuclease M20 Participates in the Down-Regulation of Mitochondrial DNA in Pollen Cells

Journal: Plant Physiology

doi: 10.1104/pp.18.00754

Generation of the atm20 mutant using CRISPR/Cas9 technology. A, Schematic diagram of the CRISPR construct containing a Cas9 expression cassette driven by an enhanced CaMV 35S promoter and an sgRNA controlled by the AtU6-26 promoter. B, A 4-bp deletion was detected in genomic DNA and cDNA of a T1 transformant (d4 mutation). The target sequence is shaded in blue, and the (PCI/PINT associated module) PAM domain is shaded in red. C, The 4-bp deletion causes a frame-shift within AtM20 and may lead to early translational termination. D, A smaller product (AtM20Δ) of AtM20 due to base deletion and early termination was detected in plants with the d4 mutation (top), and the mutation inactivates its nucleolytic activity in vitro (bottom). Arrowheads indicate the molecular products of AtM20 and AtM20Δ.
Figure Legend Snippet: Generation of the atm20 mutant using CRISPR/Cas9 technology. A, Schematic diagram of the CRISPR construct containing a Cas9 expression cassette driven by an enhanced CaMV 35S promoter and an sgRNA controlled by the AtU6-26 promoter. B, A 4-bp deletion was detected in genomic DNA and cDNA of a T1 transformant (d4 mutation). The target sequence is shaded in blue, and the (PCI/PINT associated module) PAM domain is shaded in red. C, The 4-bp deletion causes a frame-shift within AtM20 and may lead to early translational termination. D, A smaller product (AtM20Δ) of AtM20 due to base deletion and early termination was detected in plants with the d4 mutation (top), and the mutation inactivates its nucleolytic activity in vitro (bottom). Arrowheads indicate the molecular products of AtM20 and AtM20Δ.

Techniques Used: Mutagenesis, CRISPR, Construct, Expressing, Sequencing, Activity Assay, In Vitro

Related Articles

Clone Assay:

Article Title: Opsin gene duplication and diversification in the guppy, a model for sexual selection
Article Snippet: .. After reverse transcription, complementary DNA (cDNA) was cloned into pDNRlib vector using a creatorSMART cDNA construction kit (Clonetech). .. The cDNA was amplified by 18 cycles of long-distance PCR using BD Advantage 2 PCR enzyme mixture, following the manufacturer's protocols.

Amplification:

Article Title: Implication of the env Gene of the Human Endogenous Retrovirus W Family in the Expression of BDNF and DRD3 and Development of Recent-Onset Schizophrenia
Article Snippet: .. For every sample, complementary DNA (cDNA) was synthesized and amplified using the Clontech SMART PCR cDNA library construction kit (Clontech, Palo Alto, CA) as described before. .. Primers were designed according to the sequence of the HERV-W env ( , , )., For the first round of amplification, polymerase chain reaction (PCR) was performed with a pair of primers, P1 (5′-GAAGTAATCTCCCAACTCA-3′) and P2 (5′-TTTCAGCGGTTAGCAAGT-′3′).

Synthesized:

Article Title: Virus-Like Nanoparticle Vaccine Confers Protection against Toxoplasma gondii
Article Snippet: .. Complementary DNA (cDNA) was synthesized using a Prime Script 1st Strain CDNA Synthesis Kit (Takara, Japan). .. Toxoplasma gondii IMC gene was amplified by polymerase chain reaction (PCR) from cDNA with primers 5-AAA GAATTC ACCATGGGGAACACGGCGTGCTG-3 (EcoRI and XhoI underlined) and 5-TTA CTCGAG TTAGTTTCTGTCGTTGCTTGC-3 (EcoRI and XhoI underlined).

Article Title: Cloning and characterization of TaVIP2 gene from Triticum aestivum and functional analysis in Nicotiana tabacum
Article Snippet: .. Complementary DNA (cDNA) was synthesized using a Takara reverse transcription kit. .. Primers for TaVIP2 cloning, chromosome location, and subsequent identifying positive transgenic tobacco plants were designed by Primer 5 software and synthesized (Sangon, China) (see ).

Article Title: Implication of the env Gene of the Human Endogenous Retrovirus W Family in the Expression of BDNF and DRD3 and Development of Recent-Onset Schizophrenia
Article Snippet: .. For every sample, complementary DNA (cDNA) was synthesized and amplified using the Clontech SMART PCR cDNA library construction kit (Clontech, Palo Alto, CA) as described before. .. Primers were designed according to the sequence of the HERV-W env ( , , )., For the first round of amplification, polymerase chain reaction (PCR) was performed with a pair of primers, P1 (5′-GAAGTAATCTCCCAACTCA-3′) and P2 (5′-TTTCAGCGGTTAGCAAGT-′3′).

Polymerase Chain Reaction:

Article Title: Implication of the env Gene of the Human Endogenous Retrovirus W Family in the Expression of BDNF and DRD3 and Development of Recent-Onset Schizophrenia
Article Snippet: .. For every sample, complementary DNA (cDNA) was synthesized and amplified using the Clontech SMART PCR cDNA library construction kit (Clontech, Palo Alto, CA) as described before. .. Primers were designed according to the sequence of the HERV-W env ( , , )., For the first round of amplification, polymerase chain reaction (PCR) was performed with a pair of primers, P1 (5′-GAAGTAATCTCCCAACTCA-3′) and P2 (5′-TTTCAGCGGTTAGCAAGT-′3′).

cDNA Library Assay:

Article Title: Implication of the env Gene of the Human Endogenous Retrovirus W Family in the Expression of BDNF and DRD3 and Development of Recent-Onset Schizophrenia
Article Snippet: .. For every sample, complementary DNA (cDNA) was synthesized and amplified using the Clontech SMART PCR cDNA library construction kit (Clontech, Palo Alto, CA) as described before. .. Primers were designed according to the sequence of the HERV-W env ( , , )., For the first round of amplification, polymerase chain reaction (PCR) was performed with a pair of primers, P1 (5′-GAAGTAATCTCCCAACTCA-3′) and P2 (5′-TTTCAGCGGTTAGCAAGT-′3′).

Plasmid Preparation:

Article Title: Opsin gene duplication and diversification in the guppy, a model for sexual selection
Article Snippet: .. After reverse transcription, complementary DNA (cDNA) was cloned into pDNRlib vector using a creatorSMART cDNA construction kit (Clonetech). .. The cDNA was amplified by 18 cycles of long-distance PCR using BD Advantage 2 PCR enzyme mixture, following the manufacturer's protocols.

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  • 93
    TaKaRa human circakt3 cdna
    <t>circAKT3</t> expression is increased in CDDP-resistant GC cells and tissues. a Validated expression of 10 circRNAs in the tissues from 44 GC patients using RT-qPCR. b Expression levels of circAKT3 in CDDP-resistant and their matched sensitive parental cell lines (SGC7901CDDP, BGC823CDDP, SGC7901 and BGC823) normalized to GAPDH expression. c The existence of circAKT3 was validated by Sanger sequencing. The red arrow shows the “head-to-tail” splicing sites of circAKT3. d The existence of circAKT3 was validated in SGC7901CDDP and BGC823CDDP cell lines by RT-PCR. Divergent primers amplified circAKT3 in <t>cDNA</t> but not in genomic DNA (gDNA). GAPDH served as a negative control. e RNA from SGC7901CDDP and BGC823CDDP cells was treated with or without RNase R for RT-qPCR. The relative levels of circAKT3 and AKT3 mRNA were normalized to the values measured in the mock-treated cells. f Levels of small nucleolar RNA (U6, as a positive control for the nuclear fraction), GAPDH (positive control for cytoplasmic fraction), AKT3 mRNA and circRNAs from the nuclear and cytoplasmic fractions of SGC7901CDDP cells. g RNA stability of circular and linear transcripts of AKT3 and of 18S rRNA in SGC7901CDDP cells. h Representative images of RNA FISH of circAKT3 expression in SGC7901CDDP cells, which show that circAKT3 is predominantly localized to the cytoplasm. Nuclei were stained with DAPI. Scale bar, 10 μm. The results are presented as the mean ± SEM. * P
    Human Circakt3 Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human circakt3 cdna/product/TaKaRa
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human circakt3 cdna - by Bioz Stars, 2020-05
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    93
    TaKaRa cdna lines
    Cell line panel screen of drug sensitivity identifies BCL2L1 expression as a marker of insensitivity to E7107. a Top five genes whose mRNA expression positively correlated with maximum effect of cell killing (Emax) of E7107 profiled in 478 cancer cell lines. Pearson’s correlation coefficient R and p values were calculated using R package Hmisc 4.1-0. Lower Emax value indicates more robust cell killing activity. b Heatmap demonstrating the positive correlation between Emax of E7107 and BCL2L1 mRNA expression in 478 cancer cell lines. The heatmap was sorted by Emax of E7107 in cancer cell lines from high (less sensitive) to low (more sensitive). c Box plots showing the distribution of Emax (%) of E7107 in two groups of profiled cell lines defined by expression levels of each BCL2 family genes. For each BCL2 family genes, cell lines with expression level higher than the third quartile are classified into “high” and cell lines with expression level lower than the first quartile are classified into “low.” p Values were calculated using Student’s t test. d Growth-inhibitory activity of E7107 in lung cancer cell lines A549 and NCIH1568 upon doxycycline-induced <t>shRNA</t> knockdown of BCL2L1 . Left, Western blot analysis of BCLxL(encoded by BCL2L1 ) knockdown; Right, Growth curves of two cell lines measured by CellTiter-Glo. Data represent means ± SD of biological triplicates. e Growth-inhibitory activity of E7107 in lung cancer cell lines NCIH2110 and NCIH1568 upon stable <t>cDNA</t> expression of BCL2L1 (BCLxL). Left, Western blot analysis of BCLxL(encoded by BCL2L1 ) overexpression; Right, Growth curves of two cell lines measured by CellTiter-Glo. Data represent means ± SD of biological triplicates
    Cdna Lines, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna lines/product/TaKaRa
    Average 93 stars, based on 2 article reviews
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    TaKaRa inducible gfp crumbs3a cdna
    <t>Crumbs3a</t> dynamics in polarized MDCK monolayers A) Time course showing induced <t>GFP-Crb3a</t> protein levels in MDCK cells following treatment with 50 ng/ml doxycycline for 1 hour. Cell lysates were produced at the indicated times post-treatment and subjected to immunoblot with the indicated antibodies. The left lane shows uninduced expression; actin is included as a loading control. B) Time course for doxycycline-induced GFP-Crb3a mRNA levels harvested from cells treated in parallel to (A), as detected by qRT-PCR. Transcript levels are normalized to GAPDH and shown relative to uninduced control levels. The experiment was performed in triplicate; averaged levels are shown with standard deviations. C) and D) Cell surface Crb3a internalization assays using cell surface biotinylation (see Methods). Residual cell surface biotin is cleaved by treatment with MESNA prior to cell lysis to reveal internalized pool of biotinylated proteins (see schematic in C). C) Endogenous Crb3a and 3x myc-Crb3a were isolated from parental (left immunoblot) and 3x myc-Crb3a (right immunoblot) cells at indicated time points and assayed for biotinylation. D) Graph of Crb3a internalization assay using 1x myc-Crb3a and 3x myc-Crb3a cells, performed as in (C) except that the biotinylated Crb3a signal intensities were normalized to myc immunoblot signal. Averaged levels obtained from two experiments are shown with standard deviations.
    Inducible Gfp Crumbs3a Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/inducible gfp crumbs3a cdna/product/TaKaRa
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    inducible gfp crumbs3a cdna - by Bioz Stars, 2020-05
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    TaKaRa atxn8os cdna
    IRES activity of the <t>ATXN8OS</t> transcript. (A) ATXN8OS organization with promoter (open arrow), exons (open boxes) and functional splice donor sequences (GT) of D exons (D5, D4, D, D″ and D′) indicated. The CTG repeat tract is located in exon A. Transcription start site of exon D5 and exon D are represented by +1 and +801, respectively. (B) ATXN8OS RNA (NR_002717) generated from the splicing events represented by the wavy lines. The putative ORF initiated from AUG +1247 is indicated by the open boxes inside the RNA. The restriction enzymes and the cutting sites used to generate +801∼+1195 <t>cDNA</t> fragment of ATXN8OS are shown on the bottom of the cDNA. (C) The dual luciferase reporter plasmid had Renilla luciferase and firefly luciferase genes between the TK promoter and polyadenylation signal. The locations of Xba I, Xho I and Bam HI sites used for construction are shown on the top. (D) Relative luciferase activities generated by dual luciferase constructs with ECMV IRES and ATXN8OS +801∼+1195 cDNA fragment in HEK-293 and IMR-32 cells. Forty-eight hours following transfection, cells were harvested and luciferases activities were measured. IRES activity is expressed as percentages of the activity of the ECMV IRES, which was set at 100%. In addition, relative luciferase activities with ATXN8OS +801∼+953 and +953∼+1195 cDNA fragments were measured in HEK-293 cells, with IRES activity of +801∼+1195 set at 100%. Each value is the mean ± SD of three independent experiments each performed in duplicate.
    Atxn8os Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    circAKT3 expression is increased in CDDP-resistant GC cells and tissues. a Validated expression of 10 circRNAs in the tissues from 44 GC patients using RT-qPCR. b Expression levels of circAKT3 in CDDP-resistant and their matched sensitive parental cell lines (SGC7901CDDP, BGC823CDDP, SGC7901 and BGC823) normalized to GAPDH expression. c The existence of circAKT3 was validated by Sanger sequencing. The red arrow shows the “head-to-tail” splicing sites of circAKT3. d The existence of circAKT3 was validated in SGC7901CDDP and BGC823CDDP cell lines by RT-PCR. Divergent primers amplified circAKT3 in cDNA but not in genomic DNA (gDNA). GAPDH served as a negative control. e RNA from SGC7901CDDP and BGC823CDDP cells was treated with or without RNase R for RT-qPCR. The relative levels of circAKT3 and AKT3 mRNA were normalized to the values measured in the mock-treated cells. f Levels of small nucleolar RNA (U6, as a positive control for the nuclear fraction), GAPDH (positive control for cytoplasmic fraction), AKT3 mRNA and circRNAs from the nuclear and cytoplasmic fractions of SGC7901CDDP cells. g RNA stability of circular and linear transcripts of AKT3 and of 18S rRNA in SGC7901CDDP cells. h Representative images of RNA FISH of circAKT3 expression in SGC7901CDDP cells, which show that circAKT3 is predominantly localized to the cytoplasm. Nuclei were stained with DAPI. Scale bar, 10 μm. The results are presented as the mean ± SEM. * P

    Journal: Molecular Cancer

    Article Title: Circular RNA AKT3 upregulates PIK3R1 to enhance cisplatin resistance in gastric cancer via miR-198 suppression

    doi: 10.1186/s12943-019-0969-3

    Figure Lengend Snippet: circAKT3 expression is increased in CDDP-resistant GC cells and tissues. a Validated expression of 10 circRNAs in the tissues from 44 GC patients using RT-qPCR. b Expression levels of circAKT3 in CDDP-resistant and their matched sensitive parental cell lines (SGC7901CDDP, BGC823CDDP, SGC7901 and BGC823) normalized to GAPDH expression. c The existence of circAKT3 was validated by Sanger sequencing. The red arrow shows the “head-to-tail” splicing sites of circAKT3. d The existence of circAKT3 was validated in SGC7901CDDP and BGC823CDDP cell lines by RT-PCR. Divergent primers amplified circAKT3 in cDNA but not in genomic DNA (gDNA). GAPDH served as a negative control. e RNA from SGC7901CDDP and BGC823CDDP cells was treated with or without RNase R for RT-qPCR. The relative levels of circAKT3 and AKT3 mRNA were normalized to the values measured in the mock-treated cells. f Levels of small nucleolar RNA (U6, as a positive control for the nuclear fraction), GAPDH (positive control for cytoplasmic fraction), AKT3 mRNA and circRNAs from the nuclear and cytoplasmic fractions of SGC7901CDDP cells. g RNA stability of circular and linear transcripts of AKT3 and of 18S rRNA in SGC7901CDDP cells. h Representative images of RNA FISH of circAKT3 expression in SGC7901CDDP cells, which show that circAKT3 is predominantly localized to the cytoplasm. Nuclei were stained with DAPI. Scale bar, 10 μm. The results are presented as the mean ± SEM. * P

    Article Snippet: For the construction of circAKT3 overexpression plasmids, human circAKT3 cDNA was amplified using PrimerSTAR Max DNA Polymerase Mix (Takara, RR036A, Japan) and inserted into the pCD5-ciR vector (Greenseed Biotech Co, Guangzhou, China).

    Techniques: Expressing, Quantitative RT-PCR, Sequencing, Reverse Transcription Polymerase Chain Reaction, Amplification, Negative Control, Positive Control, Fluorescence In Situ Hybridization, Staining

    Cell line panel screen of drug sensitivity identifies BCL2L1 expression as a marker of insensitivity to E7107. a Top five genes whose mRNA expression positively correlated with maximum effect of cell killing (Emax) of E7107 profiled in 478 cancer cell lines. Pearson’s correlation coefficient R and p values were calculated using R package Hmisc 4.1-0. Lower Emax value indicates more robust cell killing activity. b Heatmap demonstrating the positive correlation between Emax of E7107 and BCL2L1 mRNA expression in 478 cancer cell lines. The heatmap was sorted by Emax of E7107 in cancer cell lines from high (less sensitive) to low (more sensitive). c Box plots showing the distribution of Emax (%) of E7107 in two groups of profiled cell lines defined by expression levels of each BCL2 family genes. For each BCL2 family genes, cell lines with expression level higher than the third quartile are classified into “high” and cell lines with expression level lower than the first quartile are classified into “low.” p Values were calculated using Student’s t test. d Growth-inhibitory activity of E7107 in lung cancer cell lines A549 and NCIH1568 upon doxycycline-induced shRNA knockdown of BCL2L1 . Left, Western blot analysis of BCLxL(encoded by BCL2L1 ) knockdown; Right, Growth curves of two cell lines measured by CellTiter-Glo. Data represent means ± SD of biological triplicates. e Growth-inhibitory activity of E7107 in lung cancer cell lines NCIH2110 and NCIH1568 upon stable cDNA expression of BCL2L1 (BCLxL). Left, Western blot analysis of BCLxL(encoded by BCL2L1 ) overexpression; Right, Growth curves of two cell lines measured by CellTiter-Glo. Data represent means ± SD of biological triplicates

    Journal: Nature Communications

    Article Title: Sensitivity to splicing modulation of BCL2 family genes defines cancer therapeutic strategies for splicing modulators

    doi: 10.1038/s41467-018-08150-5

    Figure Lengend Snippet: Cell line panel screen of drug sensitivity identifies BCL2L1 expression as a marker of insensitivity to E7107. a Top five genes whose mRNA expression positively correlated with maximum effect of cell killing (Emax) of E7107 profiled in 478 cancer cell lines. Pearson’s correlation coefficient R and p values were calculated using R package Hmisc 4.1-0. Lower Emax value indicates more robust cell killing activity. b Heatmap demonstrating the positive correlation between Emax of E7107 and BCL2L1 mRNA expression in 478 cancer cell lines. The heatmap was sorted by Emax of E7107 in cancer cell lines from high (less sensitive) to low (more sensitive). c Box plots showing the distribution of Emax (%) of E7107 in two groups of profiled cell lines defined by expression levels of each BCL2 family genes. For each BCL2 family genes, cell lines with expression level higher than the third quartile are classified into “high” and cell lines with expression level lower than the first quartile are classified into “low.” p Values were calculated using Student’s t test. d Growth-inhibitory activity of E7107 in lung cancer cell lines A549 and NCIH1568 upon doxycycline-induced shRNA knockdown of BCL2L1 . Left, Western blot analysis of BCLxL(encoded by BCL2L1 ) knockdown; Right, Growth curves of two cell lines measured by CellTiter-Glo. Data represent means ± SD of biological triplicates. e Growth-inhibitory activity of E7107 in lung cancer cell lines NCIH2110 and NCIH1568 upon stable cDNA expression of BCL2L1 (BCLxL). Left, Western blot analysis of BCLxL(encoded by BCL2L1 ) overexpression; Right, Growth curves of two cell lines measured by CellTiter-Glo. Data represent means ± SD of biological triplicates

    Article Snippet: Inducible shRNA and cDNA lines generated by lentiviral transduction were cultured according to the manufacturer’s instructions utilizing Tet System Approved fetal bovine serum (Clontech) rather than standard sera.

    Techniques: Expressing, Marker, Activity Assay, shRNA, Western Blot, Over Expression

    Crumbs3a dynamics in polarized MDCK monolayers A) Time course showing induced GFP-Crb3a protein levels in MDCK cells following treatment with 50 ng/ml doxycycline for 1 hour. Cell lysates were produced at the indicated times post-treatment and subjected to immunoblot with the indicated antibodies. The left lane shows uninduced expression; actin is included as a loading control. B) Time course for doxycycline-induced GFP-Crb3a mRNA levels harvested from cells treated in parallel to (A), as detected by qRT-PCR. Transcript levels are normalized to GAPDH and shown relative to uninduced control levels. The experiment was performed in triplicate; averaged levels are shown with standard deviations. C) and D) Cell surface Crb3a internalization assays using cell surface biotinylation (see Methods). Residual cell surface biotin is cleaved by treatment with MESNA prior to cell lysis to reveal internalized pool of biotinylated proteins (see schematic in C). C) Endogenous Crb3a and 3x myc-Crb3a were isolated from parental (left immunoblot) and 3x myc-Crb3a (right immunoblot) cells at indicated time points and assayed for biotinylation. D) Graph of Crb3a internalization assay using 1x myc-Crb3a and 3x myc-Crb3a cells, performed as in (C) except that the biotinylated Crb3a signal intensities were normalized to myc immunoblot signal. Averaged levels obtained from two experiments are shown with standard deviations.

    Journal: Traffic (Copenhagen, Denmark)

    Article Title: Snail Destabilizes Cell Surface Crumbs3a

    doi: 10.1111/j.1600-0854.2012.01376.x

    Figure Lengend Snippet: Crumbs3a dynamics in polarized MDCK monolayers A) Time course showing induced GFP-Crb3a protein levels in MDCK cells following treatment with 50 ng/ml doxycycline for 1 hour. Cell lysates were produced at the indicated times post-treatment and subjected to immunoblot with the indicated antibodies. The left lane shows uninduced expression; actin is included as a loading control. B) Time course for doxycycline-induced GFP-Crb3a mRNA levels harvested from cells treated in parallel to (A), as detected by qRT-PCR. Transcript levels are normalized to GAPDH and shown relative to uninduced control levels. The experiment was performed in triplicate; averaged levels are shown with standard deviations. C) and D) Cell surface Crb3a internalization assays using cell surface biotinylation (see Methods). Residual cell surface biotin is cleaved by treatment with MESNA prior to cell lysis to reveal internalized pool of biotinylated proteins (see schematic in C). C) Endogenous Crb3a and 3x myc-Crb3a were isolated from parental (left immunoblot) and 3x myc-Crb3a (right immunoblot) cells at indicated time points and assayed for biotinylation. D) Graph of Crb3a internalization assay using 1x myc-Crb3a and 3x myc-Crb3a cells, performed as in (C) except that the biotinylated Crb3a signal intensities were normalized to myc immunoblot signal. Averaged levels obtained from two experiments are shown with standard deviations.

    Article Snippet: Inducible GFP-Crumbs3a : cDNA was PCR amplified from mGFP-Crb3a ( ) and subcloned into pRetroX-Tight-Puro (Clontech).

    Techniques: Produced, Expressing, Quantitative RT-PCR, Lysis, Isolation

    IRES activity of the ATXN8OS transcript. (A) ATXN8OS organization with promoter (open arrow), exons (open boxes) and functional splice donor sequences (GT) of D exons (D5, D4, D, D″ and D′) indicated. The CTG repeat tract is located in exon A. Transcription start site of exon D5 and exon D are represented by +1 and +801, respectively. (B) ATXN8OS RNA (NR_002717) generated from the splicing events represented by the wavy lines. The putative ORF initiated from AUG +1247 is indicated by the open boxes inside the RNA. The restriction enzymes and the cutting sites used to generate +801∼+1195 cDNA fragment of ATXN8OS are shown on the bottom of the cDNA. (C) The dual luciferase reporter plasmid had Renilla luciferase and firefly luciferase genes between the TK promoter and polyadenylation signal. The locations of Xba I, Xho I and Bam HI sites used for construction are shown on the top. (D) Relative luciferase activities generated by dual luciferase constructs with ECMV IRES and ATXN8OS +801∼+1195 cDNA fragment in HEK-293 and IMR-32 cells. Forty-eight hours following transfection, cells were harvested and luciferases activities were measured. IRES activity is expressed as percentages of the activity of the ECMV IRES, which was set at 100%. In addition, relative luciferase activities with ATXN8OS +801∼+953 and +953∼+1195 cDNA fragments were measured in HEK-293 cells, with IRES activity of +801∼+1195 set at 100%. Each value is the mean ± SD of three independent experiments each performed in duplicate.

    Journal: PLoS ONE

    Article Title: Internal Ribosome Entry Segment Activity of ATXN8 Opposite Strand RNA

    doi: 10.1371/journal.pone.0073885

    Figure Lengend Snippet: IRES activity of the ATXN8OS transcript. (A) ATXN8OS organization with promoter (open arrow), exons (open boxes) and functional splice donor sequences (GT) of D exons (D5, D4, D, D″ and D′) indicated. The CTG repeat tract is located in exon A. Transcription start site of exon D5 and exon D are represented by +1 and +801, respectively. (B) ATXN8OS RNA (NR_002717) generated from the splicing events represented by the wavy lines. The putative ORF initiated from AUG +1247 is indicated by the open boxes inside the RNA. The restriction enzymes and the cutting sites used to generate +801∼+1195 cDNA fragment of ATXN8OS are shown on the bottom of the cDNA. (C) The dual luciferase reporter plasmid had Renilla luciferase and firefly luciferase genes between the TK promoter and polyadenylation signal. The locations of Xba I, Xho I and Bam HI sites used for construction are shown on the top. (D) Relative luciferase activities generated by dual luciferase constructs with ECMV IRES and ATXN8OS +801∼+1195 cDNA fragment in HEK-293 and IMR-32 cells. Forty-eight hours following transfection, cells were harvested and luciferases activities were measured. IRES activity is expressed as percentages of the activity of the ECMV IRES, which was set at 100%. In addition, relative luciferase activities with ATXN8OS +801∼+953 and +953∼+1195 cDNA fragments were measured in HEK-293 cells, with IRES activity of +801∼+1195 set at 100%. Each value is the mean ± SD of three independent experiments each performed in duplicate.

    Article Snippet: The ATXN8OS cDNA were then cloned into the Eco RI site of pEGFP-N1 (Clontech).

    Techniques: Activity Assay, Functional Assay, CTG Assay, Generated, Luciferase, Plasmid Preparation, Construct, Transfection

    Transient expression of ATXN8OS ORF-EGFP constructs in HEK-293 cells. (A) ORF-EGFP constructs. A 752-bp cDNA fragment containing exon D, C2 and portion of C1 was inserted into pEGFP-N1 MCS so that ATXN8OS ORF was fused in-frame with the EGFP gene to generate pCMV/+801. A +1∼+800 ATXN8OS fragment was inserted between CMV promoter and exon D of pCMV/+801 to generate pCMV/+1. In pATXN8OS/−114 and/−481, 114 and 481-bp ATXN8OS promoter fragments was used to replace the CMV promoter in pCMV/+1. (B) Real-time PCR quantification of ORF-EGFP RNA level relative to endogenous HPRT1 RNA. To normalize, expression level in pATXN8OS/−481 transfected cells is set as 1.0. (C) FACS analysis of EGFP fluorescence. Levels of EGFP were expressed as percentages of pIRES2-EGFP, which was set at 100%. Each value is the mean ± SD of three independent experiments each performed in duplicate.

    Journal: PLoS ONE

    Article Title: Internal Ribosome Entry Segment Activity of ATXN8 Opposite Strand RNA

    doi: 10.1371/journal.pone.0073885

    Figure Lengend Snippet: Transient expression of ATXN8OS ORF-EGFP constructs in HEK-293 cells. (A) ORF-EGFP constructs. A 752-bp cDNA fragment containing exon D, C2 and portion of C1 was inserted into pEGFP-N1 MCS so that ATXN8OS ORF was fused in-frame with the EGFP gene to generate pCMV/+801. A +1∼+800 ATXN8OS fragment was inserted between CMV promoter and exon D of pCMV/+801 to generate pCMV/+1. In pATXN8OS/−114 and/−481, 114 and 481-bp ATXN8OS promoter fragments was used to replace the CMV promoter in pCMV/+1. (B) Real-time PCR quantification of ORF-EGFP RNA level relative to endogenous HPRT1 RNA. To normalize, expression level in pATXN8OS/−481 transfected cells is set as 1.0. (C) FACS analysis of EGFP fluorescence. Levels of EGFP were expressed as percentages of pIRES2-EGFP, which was set at 100%. Each value is the mean ± SD of three independent experiments each performed in duplicate.

    Article Snippet: The ATXN8OS cDNA were then cloned into the Eco RI site of pEGFP-N1 (Clontech).

    Techniques: Expressing, Construct, Real-time Polymerase Chain Reaction, Transfection, FACS, Fluorescence