Protocol for recombinant rabies virus titration by quantitative PCR
Summary: Preparing high-titer virus and performing accurate titer determination are critical to subsequent experiments. However, not all applied recombinant rabies viruses, such as the L-deleted virus,1 are equipped with fluorescent proteins for titration by fluorescence-activated cell sorting (FACS...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-03-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007329 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832590924652216320 |
|---|---|
| author | He Zhang Xueping Gao Xiangyu Ge Xiao Wang Minghui Song Xia Zhang Lei Jin |
| author_facet | He Zhang Xueping Gao Xiangyu Ge Xiao Wang Minghui Song Xia Zhang Lei Jin |
| author_sort | He Zhang |
| collection | DOAJ |
| description | Summary: Preparing high-titer virus and performing accurate titer determination are critical to subsequent experiments. However, not all applied recombinant rabies viruses, such as the L-deleted virus,1 are equipped with fluorescent proteins for titration by fluorescence-activated cell sorting (FACS). Here, we present a quantitative reverse-transcription PCR (RT-qPCR) approach for titrating recombinant rabies virus. We describe steps for preparing standards for RT-qPCR, rabies virus genome RNA extraction, and reverse transcription of virus RNA. We then detail procedures for RT-qPCR for titration and stereotaxic rabies virus injection for titer verification. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-eb7c891d015e48cbbb3ed52a183dacf2 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-eb7c891d015e48cbbb3ed52a183dacf22025-01-23T05:27:44ZengElsevierSTAR Protocols2666-16672025-03-0161103567Protocol for recombinant rabies virus titration by quantitative PCRHe Zhang0Xueping Gao1Xiangyu Ge2Xiao Wang3Minghui Song4Xia Zhang5Lei Jin6Lingang Laboratory, Shanghai 200031, China; Corresponding authorLingang Laboratory, Shanghai 200031, ChinaLingang Laboratory, Shanghai 200031, ChinaLingang Laboratory, Shanghai 200031, ChinaLingang Laboratory, Shanghai 200031, China; School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, ChinaLingang Laboratory, Shanghai 200031, ChinaLingang Laboratory, Shanghai 200031, China; Corresponding authorSummary: Preparing high-titer virus and performing accurate titer determination are critical to subsequent experiments. However, not all applied recombinant rabies viruses, such as the L-deleted virus,1 are equipped with fluorescent proteins for titration by fluorescence-activated cell sorting (FACS). Here, we present a quantitative reverse-transcription PCR (RT-qPCR) approach for titrating recombinant rabies virus. We describe steps for preparing standards for RT-qPCR, rabies virus genome RNA extraction, and reverse transcription of virus RNA. We then detail procedures for RT-qPCR for titration and stereotaxic rabies virus injection for titer verification. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724007329Molecular BiologyNeuroscienceBiotechnology and bioengineering |
| spellingShingle | He Zhang Xueping Gao Xiangyu Ge Xiao Wang Minghui Song Xia Zhang Lei Jin Protocol for recombinant rabies virus titration by quantitative PCR STAR Protocols Molecular Biology Neuroscience Biotechnology and bioengineering |
| title | Protocol for recombinant rabies virus titration by quantitative PCR |
| title_full | Protocol for recombinant rabies virus titration by quantitative PCR |
| title_fullStr | Protocol for recombinant rabies virus titration by quantitative PCR |
| title_full_unstemmed | Protocol for recombinant rabies virus titration by quantitative PCR |
| title_short | Protocol for recombinant rabies virus titration by quantitative PCR |
| title_sort | protocol for recombinant rabies virus titration by quantitative pcr |
| topic | Molecular Biology Neuroscience Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007329 |
| work_keys_str_mv | AT hezhang protocolforrecombinantrabiesvirustitrationbyquantitativepcr AT xuepinggao protocolforrecombinantrabiesvirustitrationbyquantitativepcr AT xiangyuge protocolforrecombinantrabiesvirustitrationbyquantitativepcr AT xiaowang protocolforrecombinantrabiesvirustitrationbyquantitativepcr AT minghuisong protocolforrecombinantrabiesvirustitrationbyquantitativepcr AT xiazhang protocolforrecombinantrabiesvirustitrationbyquantitativepcr AT leijin protocolforrecombinantrabiesvirustitrationbyquantitativepcr |