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Stock No: 003605
Protocol 12209: Sanger sequencing Assay - Cln6<nclf>
Version 1.0
Notes
The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX).
To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility.
Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.
Expected Results
Sequence
ctgccctgctaaggaactcacaacttccagagggactttctatcccgtttttttttctgagggaagggttatcacacatcgtaattgaggcaggggcatgccatggccctcattcttcacctcagCTTATTGAGCGGTCCCCCC(C)GAACGCTG
CCGCGGTCTATAGTCTATGTCAGCATCATCACTTTCATCATGGGAGCCAGCATCCACCTGGTGGGAGACTCAGTCAACCATCGCCTGCTCTTCAGTGGGTACCAGC
JAX Protocol
Protocol Primers
| Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
|---|---|---|---|---|---|---|
| 20349 | CTG CCC TGC TAA GGA ACT CA | Forward | ||||
| 20350 | GCT GGT ACC CAC TGA AGA GC | Reverse |
Reaction A
| Component | Final Concentration |
|---|---|
| ddH2O | |
| High Fidelity Buffer | 1.00 |
| dNTP | 0.05 |
| MgSO4 | 1.00 |
| Platinum Taq Pol. | 0.02 |
| DNA |
Cycling
| Step | Temp °C | Time | Note |
|---|---|---|---|
| 1 | 94.0 | -- | |
| 2 | 94.0 | -- | |
| 3 | 55.0 | -- | |
| 4 | 68.0 | -- | |
| 5 | -- | Go to step 2-4, 35 times | |
| 6 | 68.0 | -- | |
| 7 | 4.0 | -- | hold |
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.
Strains Using This Protocol
This is the only strain that uses this protocol.