Stock Number:008398
Strain Name:STOCK Fxntm1Mkn Tg(FXN)YG8Pook/J
Fxntm1Pand
Fxntm1Pand GAA repeats
Standard PCR
1.0
08-June -2009 10-December -2010
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.) THIS PROTOCOL DOES NOT NEED THE MERCAPTOETHANOL ADDED.

Size on ABI 3730.
Expected Results:

JR8470, JR11113, JR12329, JR14162 in 900-950 bp range

B6 = no band
human = ~194 bp
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
8579 GAC TAA CCT GGC CAA CAT GG -
8580 6-FAM CTT GGC TTA ATG CAA CCT CTG -
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
10 X AM Buffer 1.31 1.09 X 1.31
ddH2O 6.91 - 6.91
DMSO 0.60 - 0.60
2.5 mM dNTP 1.04 0.22 mM 1.04
6.6 uM 8579 0.24 0.13 uM 0.24
6.6 uM 8580 0.24 0.13 uM 0.24
5 U/ul 5prime HotMaster Taq 0.06 0.03 U/ul 0.06
DNA (crude prep diluted 1:1 in water) 1.00 0.00 1.00
5 M Betaine 0.60 0.25 M 0.60
Cycling 
Step # Temp °C Time Note
1 - - put samples into block preheated to 94
2 94 2:00 -
3 94 0:20 -
4 59 0:40 -
5 70 1:30 -
6 - - to to step 3 additional 37 times
7 70 5:00 -
8 10 hold -
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
008398 STOCK Fxntm1Mkn Tg(FXN)YG8Pook/J
008470 B6.129-Fxntm1Pand/J
011113 B6.129-Fxntm1.1Pand/J
012329 B6.Cg-Fxntm1.1Pand Fxntm1Mkn/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.