transgene transgene junction  (Thermo Fisher)


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    Structured Review

    Thermo Fisher transgene transgene junction
    Establishment of the PgkEGFP-NOG mouse. A. A schematic representation of the inserted transgenes. The GFPF1+GFPR1 primer set was used for genotyping PCR, and the GFPF1+PGKR1 primer set was used to sequence the tandem <t>transgene</t> junction. The nucleotide sequence of the tandem transgene junction was described. B. The PCR products of the primer sets GFPF1+GFPR1 (630 bp) and GFPF1+PGKR1 (1288 bp). NOG mice were used as negative controls (Non). Tg indicates the PgkEGFP-NOG offspring. C. HCT 116 cells were transplanted into subcutaneous (a), kidney (b), testis (c), and liver tissues (d) of the PgkEGFP-NOG mice. Xenotransplanted organs were recovered 2 weeks after transplantation, and fresh frozen sections were analyzed by fluorescence microscopy. The green fluorescence indicates EGFP, and blue indicates nuclear staining (DAPI). Xenografts in the PgkEGFP-NOG mice were clearly identified as EGFP-negative colonies. *Bar=200 µ m.
    Transgene Transgene Junction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 19162 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    transgene transgene junction - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "A Novel Enhanced Green FluorescentProtein-Expressing NOG Mouse for Analyzingthe Microenvironment of Xenograft Tissues"

    Article Title: A Novel Enhanced Green FluorescentProtein-Expressing NOG Mouse for Analyzingthe Microenvironment of Xenograft Tissues

    Journal: Experimental Animals

    doi: 10.1538/expanim.63.55

    Establishment of the PgkEGFP-NOG mouse. A. A schematic representation of the inserted transgenes. The GFPF1+GFPR1 primer set was used for genotyping PCR, and the GFPF1+PGKR1 primer set was used to sequence the tandem transgene junction. The nucleotide sequence of the tandem transgene junction was described. B. The PCR products of the primer sets GFPF1+GFPR1 (630 bp) and GFPF1+PGKR1 (1288 bp). NOG mice were used as negative controls (Non). Tg indicates the PgkEGFP-NOG offspring. C. HCT 116 cells were transplanted into subcutaneous (a), kidney (b), testis (c), and liver tissues (d) of the PgkEGFP-NOG mice. Xenotransplanted organs were recovered 2 weeks after transplantation, and fresh frozen sections were analyzed by fluorescence microscopy. The green fluorescence indicates EGFP, and blue indicates nuclear staining (DAPI). Xenografts in the PgkEGFP-NOG mice were clearly identified as EGFP-negative colonies. *Bar=200 µ m.
    Figure Legend Snippet: Establishment of the PgkEGFP-NOG mouse. A. A schematic representation of the inserted transgenes. The GFPF1+GFPR1 primer set was used for genotyping PCR, and the GFPF1+PGKR1 primer set was used to sequence the tandem transgene junction. The nucleotide sequence of the tandem transgene junction was described. B. The PCR products of the primer sets GFPF1+GFPR1 (630 bp) and GFPF1+PGKR1 (1288 bp). NOG mice were used as negative controls (Non). Tg indicates the PgkEGFP-NOG offspring. C. HCT 116 cells were transplanted into subcutaneous (a), kidney (b), testis (c), and liver tissues (d) of the PgkEGFP-NOG mice. Xenotransplanted organs were recovered 2 weeks after transplantation, and fresh frozen sections were analyzed by fluorescence microscopy. The green fluorescence indicates EGFP, and blue indicates nuclear staining (DAPI). Xenografts in the PgkEGFP-NOG mice were clearly identified as EGFP-negative colonies. *Bar=200 µ m.

    Techniques Used: Polymerase Chain Reaction, Sequencing, Mouse Assay, Transplantation Assay, Fluorescence, Microscopy, Staining

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    Amplification:

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    High Throughput Screening Assay:

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    Synthesized:

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    Autoradiography:

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    Incubation:

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    Chloramphenicol Acetyltransferase Assay:

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    Hybridization:

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    High Performance Liquid Chromatography:

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    Polymerase Chain Reaction:

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    DNA Sequencing:

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    Sequencing:

    Article Title: Association Between Loss of Dp140 and Cognitive Impairment in Duchenne and Becker Dystrophies
    Article Snippet: The PCR products were purified by a PCR Product Pre-Sequencing Kit (Affymetrix Inc., Santa Clara, CA, USA), containing 4 U exonuclease I (10 U/μL) and 0.8 U shrimp alkaline phosphatase (2 U/μL). .. The sequencing reaction was performed by ABI PRISM™ BigDye Terminator v.3.1 Cycle Sequencing Kit (Applied Biosystems) and analyzed on an ABI PRISM™ 310 genetic analyzer (Applied Biosystems).

    Article Title: X-Linked Nonsyndromic Sinus Node Dysfunction and Atrial Fibrillation Caused by Emerin Mutation
    Article Snippet: For sequencing, primer pairs for PCR amplification of the six translated exons of EMD were designed using OLIGO v6.51 Primer Analysis Software (National Biosciences, Plymouth, MN, USA): EMD1F: 5′-TGCTCGGCCGGTTTTGGTAG-3′; EMD2R: 5′-CCTTTCTCCAGTGCCGCTCT-3′; EMD3F: 5′-GAGAAAGGGGAGGGAAGTCT-3′; EMD4R: 5′-GCCCAAGAGCCACCATTTGT-3′; EMD5F: 5′-GTCCCCTCGCCCTGACTCTC-3′; EMD6R: 5′-CCCCCACCCCCACTGCTAAG-3′. .. Amplified products were treated with the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, OH, USA) and sequenced by the dye-terminator method in a core facility, using an ABI PRISM 3730 XL DNA Analyzer (Applied Biosystems).

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    Article Snippet: Samples yielding anomalous traces were selected for further analysis by sequencing. .. Genomic DNA was reamplified by PCR, and products were treated with the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, OH).

    Article Title: TMEM165 Deficiency: Postnatal Changes in Glycosylation
    Article Snippet: .. After 35 cycles of 94°C for 1 min, 60°C for 1 min, and 72°C for 1.5 min, a final incubation was performed at 72°C for 10 min. PCR products were purified using the PCR Product Pre-Sequencing Kit (USB Products/Affymetrix, Ohio, USA), and sequencing was performed with the BigDye Terminator Kit 3.1 (Applied Biosystems/Life Technologies, Darmstadt, Germany). .. 1.5 μL of purified PCR product was supplemented with 1.5 μL sequencing primer (10 pmol/L), 1 μL buffer, 0.5 μL BigDye, and 5.5 μL water.

    Article Title: Development of a Melting Curve-Based Allele-Specific PCR of Apolipoprotein E (APOE) Genotyping Method for Genomic DNA, Guthrie Blood Spot, and Whole Blood
    Article Snippet: Paragraph title: Sanger sequencing for APOE genotyping ... Aliquots of PCR product were processed using a PCR Pre-Sequencing Kit (USB Corp. Cleveland, OH) to remove residual primers and dNTPs following the manufacturer's protocol.

    Article Title: Assignment of Reference 5'-end 16S rDNA Sequences and Species-Specific Sequence Polymorphisms Improves Species Identification of Nocardia
    Article Snippet: .. The cycling conditions were: 95°C for 15 min followed by 35 cycles of 94°C for 30 s, 55°C for 30 s, 72°C for 90 s with a final extension step at 72°C for 10 min. PCR products were purified (PCR Product Pre-sequencing Kit; USB Corporation, Cleveland, OH) and sequenced using the BigDye Terminator version 3.1 cycle sequencing kit (ABI PRISM 3100 genetic analyser; Applied Biosystems, Foster City, CA) and the primer 16S-27f (5’ to 3’: 3 AGA GTT TTG ATC MTG GCT CAA G 23) [ ]. ..

    Article Title: Autophagy limits the cytotoxic effects of the AKT inhibitor AZ7328 in human bladder cancer cells
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    Article Title: Association Analysis of GABRB3 Promoter Variants with Heroin Dependence
    Article Snippet: Paragraph title: Direct sequencing of the 5′ regulatory region ... PCR cycling conditions consisted of an initial denaturation at 95°C for 10 min, followed by 30 cycles of 95°C for 1 min, an annealing temperature 60°C for each amplicon for 1 min, and 72°C for 1 min. After PCR amplification, aliquots of PCR products were processed using a PCR Pre-Sequencing Kit (USB Corp., Cleveland, OH) to remove residual primers and dNTPs.

    Article Title: Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing
    Article Snippet: .. ERG11 sequence analysis PCR products were purified using the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, Ohio USA) and sequenced using ERG11-S and ERG11-A primers, and the BigDye Terminator (version 3.1) cycle sequencing kit in the ABI PRISM 3100 genetic analyser (Applied Biosystems, Foster City, CA). .. Sequences were entered into a BLASTn sequence analysis search and analyzed using editing and analyses programs in the BioManager (ANGIS) facility (accessed via. http://angis.org.au/ ).

    Article Title: Expression-independent gene trap vectors for random and targeted mutagenesis in embryonic stem cells
    Article Snippet: Paragraph title: PCR analysis and sequencing ... The resulting PCR products were cleaned up using the PCR Product Pre-Sequencing kit (USB Corporation) and sequenced directly using primers NeoC or T2 (for sequences see Supplementary Table 1 ).

    Article Title: Life with too much polyprenol-polyprenol reductase deficiency
    Article Snippet: Afterwards PCR-products were treated with the USB PCR Product Pre-Sequencing Kit from Affymetrix/USB and with the BigDye Terminator Kit 3.1 (Applied Biosystems) under the terms of the manufacturer's protocol. .. Purification was performed with Sephadex/Millipore System (Pharmacia; Millipore; Applied Biosystems) and sequencing was done on an ABI Prism 3730.

    Cellular Antioxidant Activity Assay:

    Article Title: Assignment of Reference 5'-end 16S rDNA Sequences and Species-Specific Sequence Polymorphisms Improves Species Identification of Nocardia
    Article Snippet: .. The cycling conditions were: 95°C for 15 min followed by 35 cycles of 94°C for 30 s, 55°C for 30 s, 72°C for 90 s with a final extension step at 72°C for 10 min. PCR products were purified (PCR Product Pre-sequencing Kit; USB Corporation, Cleveland, OH) and sequenced using the BigDye Terminator version 3.1 cycle sequencing kit (ABI PRISM 3100 genetic analyser; Applied Biosystems, Foster City, CA) and the primer 16S-27f (5’ to 3’: 3 AGA GTT TTG ATC MTG GCT CAA G 23) [ ]. ..

    Mutagenesis:

    Article Title: Association Between Loss of Dp140 and Cognitive Impairment in Duchenne and Becker Dystrophies
    Article Snippet: The entire coding region of the DMD gene, including exon/intron boundaries, was sequenced in patients with no detectable mutation on MLPA. .. The PCR products were purified by a PCR Product Pre-Sequencing Kit (Affymetrix Inc., Santa Clara, CA, USA), containing 4 U exonuclease I (10 U/μL) and 0.8 U shrimp alkaline phosphatase (2 U/μL).

    Article Title: TMEM165 Deficiency: Postnatal Changes in Glycosylation
    Article Snippet: Paragraph title: Mutation Analysis ... After 35 cycles of 94°C for 1 min, 60°C for 1 min, and 72°C for 1.5 min, a final incubation was performed at 72°C for 10 min. PCR products were purified using the PCR Product Pre-Sequencing Kit (USB Products/Affymetrix, Ohio, USA), and sequencing was performed with the BigDye Terminator Kit 3.1 (Applied Biosystems/Life Technologies, Darmstadt, Germany).

    Article Title: Life with too much polyprenol-polyprenol reductase deficiency
    Article Snippet: Paragraph title: 2.6. Mutation analysis of SRD5A3 ... Afterwards PCR-products were treated with the USB PCR Product Pre-Sequencing Kit from Affymetrix/USB and with the BigDye Terminator Kit 3.1 (Applied Biosystems) under the terms of the manufacturer's protocol.

    Isolation:

    Article Title: Expression-independent gene trap vectors for random and targeted mutagenesis in embryonic stem cells
    Article Snippet: The resulting PCR products were cleaned up using the PCR Product Pre-Sequencing kit (USB Corporation) and sequenced directly using primers NeoC or T2 (for sequences see Supplementary Table 1 ). .. For semi-quantitative RT–PCR analysis, RNA was isolated from wild-type E14TG2a cells as described above and cDNA synthesis was performed using an oligodT primer and Superscript III Reverse Transcriptase (Invitrogen) following the manufacturer's instructions.

    Article Title: Life with too much polyprenol-polyprenol reductase deficiency
    Article Snippet: With QIAmp DNA mini kit (QIAGEN) genomic DNA was isolated from the patient's EDTA blood sample after the manufacturer's protocol. .. Afterwards PCR-products were treated with the USB PCR Product Pre-Sequencing Kit from Affymetrix/USB and with the BigDye Terminator Kit 3.1 (Applied Biosystems) under the terms of the manufacturer's protocol.

    Purification:

    Article Title: Association Between Loss of Dp140 and Cognitive Impairment in Duchenne and Becker Dystrophies
    Article Snippet: .. The PCR products were purified by a PCR Product Pre-Sequencing Kit (Affymetrix Inc., Santa Clara, CA, USA), containing 4 U exonuclease I (10 U/μL) and 0.8 U shrimp alkaline phosphatase (2 U/μL). .. The sequencing reaction was performed by ABI PRISM™ BigDye Terminator v.3.1 Cycle Sequencing Kit (Applied Biosystems) and analyzed on an ABI PRISM™ 310 genetic analyzer (Applied Biosystems).

    Article Title: Reverse Line Blot Hybridization Assay for Identification of Medically Important Fungi from Culture and Clinical Specimens ▿
    Article Snippet: .. PCR products were purified using the PCR Product Pre-sequencing kit (USB Corporation, Cleveland, OH) and were sequenced using forward primer and the BigDye Terminator (version 3.1) cycle sequencing kit in the ABI PRISM 3100 genetic analyzer (Applied Biosystems). .. Sequences were entered into a BLASTn sequence analysis search ( ) (accessed via ).

    Article Title: TMEM165 Deficiency: Postnatal Changes in Glycosylation
    Article Snippet: .. After 35 cycles of 94°C for 1 min, 60°C for 1 min, and 72°C for 1.5 min, a final incubation was performed at 72°C for 10 min. PCR products were purified using the PCR Product Pre-Sequencing Kit (USB Products/Affymetrix, Ohio, USA), and sequencing was performed with the BigDye Terminator Kit 3.1 (Applied Biosystems/Life Technologies, Darmstadt, Germany). .. 1.5 μL of purified PCR product was supplemented with 1.5 μL sequencing primer (10 pmol/L), 1 μL buffer, 0.5 μL BigDye, and 5.5 μL water.

    Article Title: Development of a Melting Curve-Based Allele-Specific PCR of Apolipoprotein E (APOE) Genotyping Method for Genomic DNA, Guthrie Blood Spot, and Whole Blood
    Article Snippet: Aliquots of PCR product were processed using a PCR Pre-Sequencing Kit (USB Corp. Cleveland, OH) to remove residual primers and dNTPs following the manufacturer's protocol. .. The purified PCR products were subjected to Sanger sequencing using the APOE-Seq-F as the sequencing primer and an ABI PRISM® BigDye Terminator Cycle Sequencing Ready Reaction Kit Version 3.1 (Perkin Elmer Applied Biosystems, Foster City, CA), according to the manufacturer's protocol.

    Article Title: Assignment of Reference 5'-end 16S rDNA Sequences and Species-Specific Sequence Polymorphisms Improves Species Identification of Nocardia
    Article Snippet: .. The cycling conditions were: 95°C for 15 min followed by 35 cycles of 94°C for 30 s, 55°C for 30 s, 72°C for 90 s with a final extension step at 72°C for 10 min. PCR products were purified (PCR Product Pre-sequencing Kit; USB Corporation, Cleveland, OH) and sequenced using the BigDye Terminator version 3.1 cycle sequencing kit (ABI PRISM 3100 genetic analyser; Applied Biosystems, Foster City, CA) and the primer 16S-27f (5’ to 3’: 3 AGA GTT TTG ATC MTG GCT CAA G 23) [ ]. ..

    Article Title: Association Analysis of GABRB3 Promoter Variants with Heroin Dependence
    Article Snippet: PCR cycling conditions consisted of an initial denaturation at 95°C for 10 min, followed by 30 cycles of 95°C for 1 min, an annealing temperature 60°C for each amplicon for 1 min, and 72°C for 1 min. After PCR amplification, aliquots of PCR products were processed using a PCR Pre-Sequencing Kit (USB Corp., Cleveland, OH) to remove residual primers and dNTPs. .. These purified PCR products were then subjected to direct sequencing using an ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit Version 3.1, and an ABI autosequencer 3730 (Perkin Elmer Applied Biosystems, Foster City, CA), according to the manufacturer's protocol.

    Article Title: Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing
    Article Snippet: .. ERG11 sequence analysis PCR products were purified using the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, Ohio USA) and sequenced using ERG11-S and ERG11-A primers, and the BigDye Terminator (version 3.1) cycle sequencing kit in the ABI PRISM 3100 genetic analyser (Applied Biosystems, Foster City, CA). .. Sequences were entered into a BLASTn sequence analysis search and analyzed using editing and analyses programs in the BioManager (ANGIS) facility (accessed via. http://angis.org.au/ ).

    Article Title: Life with too much polyprenol-polyprenol reductase deficiency
    Article Snippet: Afterwards PCR-products were treated with the USB PCR Product Pre-Sequencing Kit from Affymetrix/USB and with the BigDye Terminator Kit 3.1 (Applied Biosystems) under the terms of the manufacturer's protocol. .. Purification was performed with Sephadex/Millipore System (Pharmacia; Millipore; Applied Biosystems) and sequencing was done on an ABI Prism 3730.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Expression-independent gene trap vectors for random and targeted mutagenesis in embryonic stem cells
    Article Snippet: The resulting PCR products were cleaned up using the PCR Product Pre-Sequencing kit (USB Corporation) and sequenced directly using primers NeoC or T2 (for sequences see Supplementary Table 1 ). .. For semi-quantitative RT–PCR analysis, RNA was isolated from wild-type E14TG2a cells as described above and cDNA synthesis was performed using an oligodT primer and Superscript III Reverse Transcriptase (Invitrogen) following the manufacturer's instructions.

    Polyacrylamide Gel Electrophoresis:

    Article Title: X-Linked Nonsyndromic Sinus Node Dysfunction and Atrial Fibrillation Caused by Emerin Mutation
    Article Snippet: Genotyping was accomplished by PCR amplification of genomic DNA radio-labeled with alpha-(32 P)-dCTP, resolution of alleles by polyacrylamide gel electrophoresis, and visualization by autoradiography, as previously described. .. Amplified products were treated with the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, OH, USA) and sequenced by the dye-terminator method in a core facility, using an ABI PRISM 3730 XL DNA Analyzer (Applied Biosystems).

    Multiplex Ligation-dependent Probe Amplification:

    Article Title: Association Between Loss of Dp140 and Cognitive Impairment in Duchenne and Becker Dystrophies
    Article Snippet: The entire coding region of the DMD gene, including exon/intron boundaries, was sequenced in patients with no detectable mutation on MLPA. .. The PCR products were purified by a PCR Product Pre-Sequencing Kit (Affymetrix Inc., Santa Clara, CA, USA), containing 4 U exonuclease I (10 U/μL) and 0.8 U shrimp alkaline phosphatase (2 U/μL).

    Activated Clotting Time Assay:

    Article Title: Life with too much polyprenol-polyprenol reductase deficiency
    Article Snippet: For amplification the following primers synthesized by Invitrogen were used: SRD5A3 IF(13U): (5′ to 3′) TGT AAA ACG ACG GCC AGT AGG CTG AGA CCG GTG CGC CG; SRD5A3 IR(13R): (5′ to 3′) CAG GAA ACA GCT ATG ACC GCC ATC CAT TGG CAC TTG GC; SRD5A3 IIF (13U): (5′ to 3′) TGT AAA ACG ACG GCC AGT CCT GCT TTG GTG CCT TAC TC; SRD5A3 IIR(13R): (5′ to 3′) CAG GAA ACA GCT ATG ACC ACT GCC ATG CTC ATT CAG TG. .. Afterwards PCR-products were treated with the USB PCR Product Pre-Sequencing Kit from Affymetrix/USB and with the BigDye Terminator Kit 3.1 (Applied Biosystems) under the terms of the manufacturer's protocol.

    Software:

    Article Title: Association Between Loss of Dp140 and Cognitive Impairment in Duchenne and Becker Dystrophies
    Article Snippet: The MLPA data interpretation, in order to assess copy number changes (deletions) in comparison to the normal controls, was performed by the Excel program by Coffalyser MLPA data analysis software [ www.mlpa.con ]. .. The PCR products were purified by a PCR Product Pre-Sequencing Kit (Affymetrix Inc., Santa Clara, CA, USA), containing 4 U exonuclease I (10 U/μL) and 0.8 U shrimp alkaline phosphatase (2 U/μL).

    Article Title: X-Linked Nonsyndromic Sinus Node Dysfunction and Atrial Fibrillation Caused by Emerin Mutation
    Article Snippet: For sequencing, primer pairs for PCR amplification of the six translated exons of EMD were designed using OLIGO v6.51 Primer Analysis Software (National Biosciences, Plymouth, MN, USA): EMD1F: 5′-TGCTCGGCCGGTTTTGGTAG-3′; EMD2R: 5′-CCTTTCTCCAGTGCCGCTCT-3′; EMD3F: 5′-GAGAAAGGGGAGGGAAGTCT-3′; EMD4R: 5′-GCCCAAGAGCCACCATTTGT-3′; EMD5F: 5′-GTCCCCTCGCCCTGACTCTC-3′; EMD6R: 5′-CCCCCACCCCCACTGCTAAG-3′. .. Amplified products were treated with the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, OH, USA) and sequenced by the dye-terminator method in a core facility, using an ABI PRISM 3730 XL DNA Analyzer (Applied Biosystems).

    Article Title: A Common Polymorphism in SCN5A is Associated with Lone Atrial Fibrillation
    Article Snippet: Primer pairs for exon-specific PCR amplification of genomic DNA were designed using OLIGO v6.51 Primer Analysis Software (National Biosciences, Plymouth, MN) and WAVEMAKER version 4.0.32 software (Transgenomic, Omaha, NE). .. Genomic DNA was reamplified by PCR, and products were treated with the PCR Product Pre-sequencing Kit (USB Corporation, Cleveland, OH).

    Article Title: TMEM165 Deficiency: Postnatal Changes in Glycosylation
    Article Snippet: Primers for sequencing of TMEM165 genomic DNA , designed with Primer3 software (Rozen and Skaletsky ) and purchased from Invitrogen (Carlsbad, CA, USA), are listed in the online supplementary material (see Table 1S). .. After 35 cycles of 94°C for 1 min, 60°C for 1 min, and 72°C for 1.5 min, a final incubation was performed at 72°C for 10 min. PCR products were purified using the PCR Product Pre-Sequencing Kit (USB Products/Affymetrix, Ohio, USA), and sequencing was performed with the BigDye Terminator Kit 3.1 (Applied Biosystems/Life Technologies, Darmstadt, Germany).

    Article Title: Autophagy limits the cytotoxic effects of the AKT inhibitor AZ7328 in human bladder cancer cells
    Article Snippet: Five nanograms of genomic DNA from each cell line were amplified in an 8-L PCR using AmpliTaq Gold (Applied Biosystems) on PE 9700 machines and subsequently cleaned using a diluted version of the EXO-SAP-based PCR product pre-sequencing kit (USB Corp.) dispensed by a nanoliter dispenser (Deerac Fluidics Equator by Promega). .. Base calling, quality assessment and assembly were performed using the Phred, Phrap, Polyphred, Consed software suite.

    Multiplex Assay:

    Article Title: Reverse Line Blot Hybridization Assay for Identification of Medically Important Fungi from Culture and Clinical Specimens ▿
    Article Snippet: For fungal strains, the ITS1, 5.8S, and ITS2 regions were amplified using the SR6RL and LR1L primer pair as described for multiplex PCR (see above). .. PCR products were purified using the PCR Product Pre-sequencing kit (USB Corporation, Cleveland, OH) and were sequenced using forward primer and the BigDye Terminator (version 3.1) cycle sequencing kit in the ABI PRISM 3100 genetic analyzer (Applied Biosystems).

    CTG Assay:

    Article Title: Life with too much polyprenol-polyprenol reductase deficiency
    Article Snippet: For amplification the following primers synthesized by Invitrogen were used: SRD5A3 IF(13U): (5′ to 3′) TGT AAA ACG ACG GCC AGT AGG CTG AGA CCG GTG CGC CG; SRD5A3 IR(13R): (5′ to 3′) CAG GAA ACA GCT ATG ACC GCC ATC CAT TGG CAC TTG GC; SRD5A3 IIF (13U): (5′ to 3′) TGT AAA ACG ACG GCC AGT CCT GCT TTG GTG CCT TAC TC; SRD5A3 IIR(13R): (5′ to 3′) CAG GAA ACA GCT ATG ACC ACT GCC ATG CTC ATT CAG TG. .. Afterwards PCR-products were treated with the USB PCR Product Pre-Sequencing Kit from Affymetrix/USB and with the BigDye Terminator Kit 3.1 (Applied Biosystems) under the terms of the manufacturer's protocol.

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