All loci were co-amplified using HuaXia Platinum PCR Amplification kit in GeneAmp 9700 PCR system (Applied Biosystems, Foster City, USA) under the producer’s specification. Thermal cycler conditions were as described below: pre-denaturation at 95°C for 1 min, followed by 27 cycles of 94°C for 3 s, 59°C for 16 s, 65°C for 29 s, and a final extension for 5 min at 60°C. The whole PCR reaction could be finished in 1 hour. Separation of amplified products was performed by capillary electrophoresis on the ABI 3130xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) following the instructions in 11 µl reactions which consist of 1 µl PCR product or allelic ladder and the mixture of 9.6 µl Hi-Di Formamide and 0.4 µl GeneScan®-600 LIZ® Size Standard v2.0. The loading mixture was first denatured at 95°C for 3 min, followed by cooling at 4°C for 3 min immediately. The results of STRs typing were identified by the GeneMapper ID 3.2 software (Applied Biosystems, Foster City, CA, USA). Deionized water and control DNA from human cell line 9948 were typed as negative and positive control, respectively.
Genemapper id 3
Manufactured by Thermo Fisher Scientific
Sourced in United States
GeneMapper ID 3.2 software is a data analysis software for DNA fragment analysis. It is designed to automate the analysis of DNA fragments generated using capillary electrophoresis instruments. The software processes raw data and generates results, including fragment sizes and peak information.
Automatically generated - may contain errors
Lab products found in correlation
Geneamp pcr system 9700 thermal cycler, by Thermo Fisher Scientific (3 mentions)
Genescan 600 liz size standard v2, by Thermo Fisher Scientific (1 mentions)
Abi 3130xl genetic analyzer, by Thermo Fisher Scientific (1 mentions)
Geneamp pcr system 9700, by Thermo Fisher Scientific (1 mentions)
Abi 3500, by Thermo Fisher Scientific (1 mentions)
Abi prism 3130 genetic analyzer, by Thermo Fisher Scientific (1 mentions)
Abi prism 3130xl genetic analyzer, by Thermo Fisher Scientific (1 mentions)
Powerplex 21 system, by Promega (1 mentions)
Human umbilical vein endothelial cells (huvec), by Cyagen (1 mentions)
6 protocols using genemapper id 3
1
Multiplex PCR and Capillary Electrophoresis for STR Typing
2
Y-STR Profiling by Capillary Electrophoresis
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Y‐STR amplification was performed on a GeneAmp® PCR System 9700 Thermal Cycler (Applied Biosystems) using the Yfiler® Plus PCR Amplification kit according to the manufacturer's instruction. Amplified products were separated and detected by capillary electrophoresis in ABI 3500 (Applied Biosystems). The Y‐STR genotypes were analyzed using the GeneMapper ID 3.2 software (Applied Biosystems).
3
Multiplex PCR of 21 STR Loci
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The 21 non-CODIS STR loci were amplified simultaneously in a single polymerase chain reaction (PCR) system by the AGCU 21+1 fluorescence amplification reagents (AGCU ScienTech Incorporation, Wuxi, Jiangsu, China) with a GeneAmp PCR System 9700 Thermal Cycler (Applied Biosystems, Foster City, CA, USA). The PCR conditions could be obtained from the developmental validation of the reagent [9 (link)]. STR genotyping was performed on an ABI PRISM 3130 Genetic Analyzer (Applied Biosystems) and analysed by the Genemapper ID 3.2 software (Applied Biosystems).
4
Multi-Dimensional Analysis of Samples
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The software used for data collection in this study includes Malvern: Zetasizer Software 7.01, Microplate reader: SkanIt 2.4.3.37 and Confocal laser scanning microscopy: NIS 4.13. The software used for data analysis in this study includes GraphPad prism 8.0, Microsoft Office 2019, ImageJ 1.8.0, FlowJo_V10, Trim-galore (version 0.6.7); HISAT2 (version 2.2.1), featureCounts (version 2.0.1), DESeq2 (version 1.32.0) and R package clusterProfiler, ABI Prism 3130 XL Genetic Analyzer, Gene Mapper ID 3.2 software (Applied Biosystems).
5
PCR Amplification Using PowerPlex 21 System
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PCR amplification was performed using the PowerPlex® 21 System (Promega, Madison, WI, USA). The total volume of PCR reactions was 25 µL, containing 5 µL PowerPlex® 21 5× Master Mix, 5 µL PowerPlex® 21 5× Primer Pair Mix, 1 ng template DNA and amplification grade water. Amplification was carried out using a GeneAmp PCR System 9700 Thermal Cycler (Applied Biosystems, Foster City, CA, USA) under the manufacturer’s specifications. The AB PRISM 3130 Genetic Analyzer (Applied Biosystems) was used to obtain sample genotypes. Raw data were analysed using GeneMapper ID 3.2 software (Applied Biosystems). 9947A DNA was used as a positive control.
6
Characterization of HUVEC and Rat/Mouse Models
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HUVEC was obtained from Cyagen Biosciences. Biotechnology Development Co., Ltd HUVEC lines validation using short tandem repeat (STR) markers were performed by Shanghai Qingqi Biotechnology Development Co., Ltd. Sample DNA was extracted using Axygen’s genome extraction kit and amplified using a 10-STR amplification protocol (D4S2408 as the human locus). STR loci were detected on an ABI model 3730XL genetic analyzer. Data were analyzed using Gene Mapper ID 3.2 software (Applied Biosystems). Appropriate positive and negative controls were run and confirmed for each sample submitted.
SD rats (male, 6 weeks old) and Kunming (KM) mice (female, 11 weeks old) were utilized and conformed to the Animal Laboratory Ethics Committee of Shenyang Pharmaceutical University. The living environment of animals was maintained at a temperature of ~25 °C with a 12 h light/dark cycle, with free access to standard food and water. All the animal experiments were conducted according to the Guidelines for the Care and Use of Laboratory Animals approved by the Institutional Animal Ethical Care Committee (IAEC) of Shenyang Pharmaceutical University.
SD rats (male, 6 weeks old) and Kunming (KM) mice (female, 11 weeks old) were utilized and conformed to the Animal Laboratory Ethics Committee of Shenyang Pharmaceutical University. The living environment of animals was maintained at a temperature of ~25 °C with a 12 h light/dark cycle, with free access to standard food and water. All the animal experiments were conducted according to the Guidelines for the Care and Use of Laboratory Animals approved by the Institutional Animal Ethical Care Committee (IAEC) of Shenyang Pharmaceutical University.
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