Stock Number:008827
Strain Name:B6.Cg-Tg(Prdm1-cre)1Masu/J

Tg(Prdm1-cre)1Masu
Tg(Prdm1-cre)
Melt Curve Analysis
2.0
28-March -2012 28-March -2012
Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals.
Expected Results:
WT - TM 81 +/- 1 Area - 8
TG/0 - TG - TM 86 +/- 1 Area - 8
            WT - TM 81 +/- 1 Area - 3
 

Transgene = 200bp
Internal positive control = 324bp

Gel Image 
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
8076 - GCC GAG GTG CGC GTC AGT AC - -
8077 - CTG AAC ATG TCC ATC AGG TTC TTG - -
oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - -
oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - -
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration
Total Volume (µl)
ddH2O 3.35 - 3.35
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
25 mM MgCl2 0.96 2.00 mM 0.96
10 mM dNTP KAPA 0.24 0.20 mM 0.24
20 uM oIMR7338 0.30 0.25 uM 0.30
20 uM oIMR7339 0.30 0.25 uM 0.30
20 uM 8076 0.30 0.25 uM 0.30
20 uM 8077 0.30 0.25 uM 0.30
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05
20 X EvaGreen 0.60 1.00 X 0.60
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 2 min -
2 94 20sec -
3 65 15sec -1.5 C per cycle
4 68 10sec -
5 - - repeat steps 2-4 for 10 cycles
6 94 15sec -
7 50 15sec -
8 72 10sec -
9 - - repeat steps 6-8 for 28 cycles
10 72 2 min -
11 10 - hold
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
008827 B6.Cg-Tg(Prdm1-cre)1Masu/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.