Validation of Using Multiplex PCR with Sex Markers SSM4 and ALLWSex2 in Long-Term Stored Blood Samples to Determine Sex of the North American Shortnose Sturgeon (Acipenser brevirostrum)
Bibliographic record
Abstract
Sex-specific information is crucial for sturgeon culture, conservation, and fisheries management. However, identifying their sex is difficult outside the spawning season. Two recently identified female-specific loci (AllWSex2 and SSM4) are conserved across many Acipenserid species, but they have not been validated for all species within this family. This study aimed to (1) determine whether SSM4 can be used to sex shortnose sturgeon, (2) develop and test a multiplex PCR technique using both ALLWSex2 and SSM4 for sexing shortnose sturgeon, (3) determine if long-term stored blood samples can be used to sex shortnose sturgeon, and (4) test the effect of storage temperature on DNA degradation. DNA was extracted from frozen RBC samples from 36 previously sexed fish. A multiplex PCR was set up using three pairs of primers: AllWSex2 and SSM4, as female-specific loci, and mtDNA, as an internal control. AllWSex2 and SSM4 allowed for perfect discrimination of sex. While long-term storage and storage temperature did cause DNA degradation, the signal was still strong enough after 8 years of cold storage for reliable sex determination. This suggests that researchers now have the ability to re-examine archived/frozen samples to determine the sex of their sturgeon.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".