Stock Number: 002809 Strain Name: B6CBA-Tg(HDexon1)61Gpb/1J

Allele: Tg(HDexon1)61Gpb Protocol Name: TG(HDexon1), TG(YAC) Method: Standard PCR Version: 1.1 Created: 05-January -2001 Updated: 26-February -2013

Notes: AM Bufer:  670 mM TrisHCl pH 8.8, 166 mM (NH4)2SO4, 20 mM MgCl2, 1.7 mg/ml BSA, 10 mM 2-mercaptoethanol (Add 2-mercaptoethanol to 10 mM just prior to use. Do not store with 2-mercaptoethanol in buffer.) Expected Results: Transgene (Reaction A) = primers flank CAG repeat so size will vary depending on CAG repeat number ({CAG repeat number x 3} + 221bp) Transgene (Reaction B) = 170 bp (primers do not flank CAG repeat)

Gel Image

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Gel Information: Separated by gel electrophoresis on a 2.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1594 - CCG CTC AGG TTC TGC TTT TA - Transgene Forward oIMR1596 - TGG AAG GAC TTG AGG GAC TC - Transgene Reverse oIMR6533 - GGC TGA GGA AGC TGA GGA G - Transgene Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 5.14 - 5.14 10 X AM Buffer 1.20 1.09 X 1.20 25 mM MgCl2 0.96 2.18 mM 0.96 2.5 mM dNTP 0.96 0.22 mM 0.96 20 uM oIMR1594 0.24 0.44 uM 0.24 20 uM oIMR6533 0.24 0.44 uM 0.24 DMSO 1.20 - 1.20 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 1.00 - 1.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 58 1 min - 4 72 3 min repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold**

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 5.14 - 5.14 10 X AM Buffer 1.20 1.09 X 1.20 25 mM MgCl2 0.96 2.18 mM 0.96 2.5 mM dNTP 0.96 0.22 mM 0.96 20 uM oIMR1594 0.24 0.44 uM 0.24 20 uM oIMR1596 0.24 0.44 uM 0.24 DMSO 1.20 - 1.20 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 1.00 - 1.00 This genotyping protocol is used for the following strains:   Stock Number Strain Name 002809 B6CBA-Tg(HDexon1)61Gpb/1J 002810 B6CBA-Tg(HDexon1)62Gpb/1J 004601 B6CBA-Tg(HDexon1)62Gpb/2J 006494 B6CBA-Tg(HDexon1)62Gpb/3J 007578 CBy.Cg-Tg(HDexon1)61Gpb/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 55 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold**