Rectal swab DNA collection protocol for PCR genotyping in rats
DNA collection is essential for genotyping laboratory animals. Common collection methods require tissue amputation, causing discomfort and injury. Rectal swabbing has been proposed as an effective, minimally invasive alternative, but an evidence-backed protocol for the technique remains unavailable....
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2024-06-01
|
| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.2144/btn-2024-0023 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841526683940683776 |
|---|---|
| author | Audrey E Kaye Jacob W Proctor-Bonbright Jai Y Yu |
| author_facet | Audrey E Kaye Jacob W Proctor-Bonbright Jai Y Yu |
| author_sort | Audrey E Kaye |
| collection | DOAJ |
| description | DNA collection is essential for genotyping laboratory animals. Common collection methods require tissue amputation, causing discomfort and injury. Rectal swabbing has been proposed as an effective, minimally invasive alternative, but an evidence-backed protocol for the technique remains unavailable. This report evaluates the effects of collection parameters on the quality of PCR results and presents a protocol for genotyping a litter of rats within 3–5 h. Samples with 2–8 scrapes produced enough DNA to amplify targets up to ∼1800 bp long using PCR. Rectal swabbing produced PCR results with similar utility as ear clip samples, and results were unaffected by residual fecal matter or cell debris. The protocol enables fast, minimally invasive and repeatable genotyping using commercial PCR reagents. |
| format | Article |
| id | doaj-art-d8d091ac6e594a1b9cbeffe5fb9ec3d2 |
| institution | Kabale University |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2024-06-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-d8d091ac6e594a1b9cbeffe5fb9ec3d22025-01-16T15:38:47ZengTaylor & Francis GroupBioTechniques0736-62051940-98182024-06-0176627528310.2144/btn-2024-0023Rectal swab DNA collection protocol for PCR genotyping in ratsAudrey E Kaye0Jacob W Proctor-Bonbright1Jai Y Yu2Neuroscience Institute, University of Chicago, Chicago, IL 60637, USANeuroscience Institute, University of Chicago, Chicago, IL 60637, USANeuroscience Institute, University of Chicago, Chicago, IL 60637, USADNA collection is essential for genotyping laboratory animals. Common collection methods require tissue amputation, causing discomfort and injury. Rectal swabbing has been proposed as an effective, minimally invasive alternative, but an evidence-backed protocol for the technique remains unavailable. This report evaluates the effects of collection parameters on the quality of PCR results and presents a protocol for genotyping a litter of rats within 3–5 h. Samples with 2–8 scrapes produced enough DNA to amplify targets up to ∼1800 bp long using PCR. Rectal swabbing produced PCR results with similar utility as ear clip samples, and results were unaffected by residual fecal matter or cell debris. The protocol enables fast, minimally invasive and repeatable genotyping using commercial PCR reagents.https://www.tandfonline.com/doi/10.2144/btn-2024-0023DNA collectionDNA collection protocolgenotypingminimally invasivePCR analysispolymerase chain reactions |
| spellingShingle | Audrey E Kaye Jacob W Proctor-Bonbright Jai Y Yu Rectal swab DNA collection protocol for PCR genotyping in rats BioTechniques DNA collection DNA collection protocol genotyping minimally invasive PCR analysis polymerase chain reactions |
| title | Rectal swab DNA collection protocol for PCR genotyping in rats |
| title_full | Rectal swab DNA collection protocol for PCR genotyping in rats |
| title_fullStr | Rectal swab DNA collection protocol for PCR genotyping in rats |
| title_full_unstemmed | Rectal swab DNA collection protocol for PCR genotyping in rats |
| title_short | Rectal swab DNA collection protocol for PCR genotyping in rats |
| title_sort | rectal swab dna collection protocol for pcr genotyping in rats |
| topic | DNA collection DNA collection protocol genotyping minimally invasive PCR analysis polymerase chain reactions |
| url | https://www.tandfonline.com/doi/10.2144/btn-2024-0023 |
| work_keys_str_mv | AT audreyekaye rectalswabdnacollectionprotocolforpcrgenotypinginrats AT jacobwproctorbonbright rectalswabdnacollectionprotocolforpcrgenotypinginrats AT jaiyyu rectalswabdnacollectionprotocolforpcrgenotypinginrats |