PCR Record ------ROSA Tg mice (ROSA)

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Standards   ROSA w, ROSA@ƒw¤@ROSA @ƒz    

Sample               

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DNA

 

 

 

(n+1)

 

2 ƒÊl

PCR

IMR314 (20ƒÊM)

0.4

ƒÊl

~

@

=          ƒÊl

 

94Ž@5min

IMR315 (20ƒÊM)

0.4

ƒÊl

~

 

=          ƒÊl

94Ž@1min

55Ž@2 min

72Ž@1min

30

cycles

IMR316 (20ƒÊM)

0.4

ƒÊl

~

 

=          ƒÊl

25mM MgCl2

2

ƒÊl

~

 

=          ƒÊl

  18 ƒÊl

10x PCR buffer

 (15 mM Mg“ü‚è)

2

ƒÊl

~

 

=          ƒÊl

‚ÂÂ

dNTP mix

(2.5 mM)

1.6

ƒÊl

~

 

=          ƒÊl

ƒ`ƒ…[ƒu‚Ö

Taq

0.1

ƒÊl

~

 

=          ƒÊl

 

DW

11.1

ƒÊl

~

 

=          ƒÊl

 

72Ž@10min

‡Œv

 

 

 

 

20 ƒÊl

4Ž@  

Gel       Agarose  1, 1.5 %

 

Marker     500 bp ladder

PCRŽY•¨@+@ Dye 3ƒÊl

 

 

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wildtype ------- 350  bp

hetero------        350 +1100

homo ------        1100 

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