Difference between revisions of "Mimulus:Microsatellite recipes"

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Line 2: Line 2:
 
*Check out the [http://mimulusevolution.org Mimulus Biological Resource Center] for map location of these microsatellies.   
 
*Check out the [http://mimulusevolution.org Mimulus Biological Resource Center] for map location of these microsatellies.   
 
*The primers for these microsatellites are archived in genbank.
 
*The primers for these microsatellites are archived in genbank.
 +
*See the [[Full Microsatellite Sequences]].
 
*In addition to these recipes, check out the [[PCR Program for Microsatellites]].   
 
*In addition to these recipes, check out the [[PCR Program for Microsatellites]].   
 
*Contact David Lowry (dbl2@duke.edu) or other members of the [[Mimulus Community]] for more inforamtion about these markers.
 
*Contact David Lowry (dbl2@duke.edu) or other members of the [[Mimulus Community]] for more inforamtion about these markers.
 +
*[[Media:Microsatellite_PCR_conditions.xls|Microsat conditions spreadsheet]]
  
Below are the recipes that have been fine-tuned for ''Mimulus guttatus'':
+
Below are the recipes that have been fine-tuned for ''Mimulus guttatus''
  
 
==AAT 9==
 
==AAT 9==
Line 14: Line 16:
 
Cycles: 26
 
Cycles: 26
  
*H20 5.05μl
+
*H20 8.05μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 1.2μl
 
*MgCl2 (25mM) 1.2μl
Line 20: Line 22:
 
*AAT9R 0.6μl
 
*AAT9R 0.6μl
 
*AAT9F 0.25μl
 
*AAT9F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
  
 
==AAT 39==
 
==AAT 39==
Line 62: Line 63:
  
  
*H20 5.05μl
+
*H20 8.05μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 1.2μl
 
*MgCl2 (25mM) 1.2μl
Line 68: Line 69:
 
*217R 0.6μl
 
*217R 0.6μl
 
*217F 0.25μl
 
*217F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
  
 
==AAT 222==
 
==AAT 222==
Line 92: Line 92:
 
Cycles: 32
 
Cycles: 32
  
*H20 5.05μl
+
*H20 8.05μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 1.2μl
 
*MgCl2 (25mM) 1.2μl
Line 98: Line 98:
 
*225R 0.6μl
 
*225R 0.6μl
 
*225F 0.25μl
 
*225F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
 
 
  
 
==AAT 230==  
 
==AAT 230==  
Line 110: Line 108:
  
 
 
*H20 3.85μl
+
*H20 6.85μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 2.4μl
 
*MgCl2 (25mM) 2.4μl
Line 116: Line 114:
 
*230R 0.6μl
 
*230R 0.6μl
 
*230F 0.25μl
 
*230F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
  
 
==AAT 233==  
 
==AAT 233==  
Line 141: Line 138:
 
Cycles: 25
 
Cycles: 25
  
*H20 4.45μl
+
*H20 7.45μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 1.8μl
 
*MgCl2 (25mM) 1.8μl
Line 147: Line 144:
 
*240R 0.6μl
 
*240R 0.6μl
 
*240F 0.25μl
 
*240F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
  
 
==AAT 242==
 
==AAT 242==
Line 206: Line 202:
  
  
Annealing Temp 49
+
Annealing Temp: 49
 +
 
 +
Cycles: 30
  
*H20 3.85μl
+
*H20 6.85μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 2.4μl
 
*MgCl2 (25mM) 2.4μl
Line 214: Line 212:
 
*267R 0.6μl
 
*267R 0.6μl
 
*267F 0.25μl
 
*267F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
  
 
==AAT 272==
 
==AAT 272==
Line 238: Line 235:
  
 
Annealing Temp: 53
 
Annealing Temp: 53
 +
 
Cycles: 24
 
Cycles: 24
  
Line 243: Line 241:
  
 
 
*H20 2.95μl
+
*H20 5.95μl
 
*10X Buffer 1.5μl
 
*10X Buffer 1.5μl
 
*MgCl2 (25mM) 1.8μl
 
*MgCl2 (25mM) 1.8μl
Line 250: Line 248:
 
*278R 0.6μl
 
*278R 0.6μl
 
*278F 0.25μl
 
*278F 0.25μl
*Taq 0.2μl
+
*Taq 0.2μl
 
 
  
 
==AAT 283==
 
==AAT 283==
Line 269: Line 266:
  
  
 +
==AAT 296==
 +
Dye: NED
  
 +
Annealing Temp: 53
  
 +
Cycles: 28
  
  
 +
*H20 2.1μl
 +
*10X Buffer 1.5μl
 +
*MgCl2 (25mM) 1.8μl
 +
*KCl (.5M) 1.5μl
 +
*dNTPs (.25mM) 1.2μl
 +
*296R 1.2μl
 +
*296F 0.5μl
 +
*Taq 0.2μl
  
  
 +
==AAT 300==
 +
Dye: FAM
  
 +
Annealing Temp 55
  
 +
Cycles 22
  
 +
 +
*H20 8.05μl
 +
*10X Buffer 1.5μl
 +
*MgCl2 (25mM) 1.2μl
 +
*dNTPs (.25mM) 1.2μl
 +
*300R 0.6μl
 +
*300F 0.25μl
 +
*Taq 0.2μl
  
 +
==AAT 308==
 +
Dye: VIC
  
 +
Annealing Temp: 55
  
 +
Cycles: 24
  
 +
*H20 5.05μl
 +
*10X Buffer 1.5μl
 +
*MgCl2 (25mM) 1.2μl
 +
*dNTPs (.25mM) 1.2μl
 +
*308R 0.6μl
 +
*308F 0.25μl
 +
*Taq 0.2μl
  
  
 +
==AAT 312==
 +
Dye: NED
  
 +
Annealing Temp: 51
  
AAT 296 - NED
+
Cycles: 30
 +
 +
*H20 2.95μl
 +
*10X Buffer 1.5μl
 +
*MgCl2 (25mM) 1.8μl
 +
*KCl (0.5M) 1.5μl
 +
*dNTPs (.25mM) 1.2μl
 +
*312R 0.6μl
 +
*312F 0.25μl
 +
*Taq 0.2μl
  
  
  
• Annealing Temp 53
+
==AAT 333==
• Cycles 28
+
Dye: VIC
  
 +
Annealing Temp: 51
  
 +
Cycles: 30
  
    per sample x15 x110
+
*H20 3.85μl
H20 2.1μl 31.5μl 231μl
+
*10X Buffer 1.5μl
10X Buffer 1.5μl 22.5μl 165μl
+
*MgCl2 (25mM) 2.4μl
MgCl2 (25mM) 1.8μl 27μl 198μl
+
*dNTPs (.25mM) 1.2μl
• KCl (.5M) 1.5μl 22.5μl 165μl
+
*333R 0.6μl
dNTPs (.25mM) 1.2μl 18μl 132μl
+
*Taq 0.2μl
• 296R ** 1.2μl 18μl 132μl **
 
• 296F ** 0.5μl 7.5μl 55μl **
 
Taq 0.2μl 3μl 22μl
 
  
  
 +
==AAT 350==
 +
Dye: FAM
  
 +
Annealing Temp: 52
  
 +
Cycles: 28
  
 +
*H20 5.05μl
 +
*10X Buffer 1.5μl
 +
*MgCl2 (25mM) 1.2μl
 +
*dNTPs (.25mM) 1.2μl
 +
*350R 0.6μl
 +
*350F 0.25μl
 +
*Taq 0.2μl
  
  
  
 +
==AAT 356==
 +
Dye: VIC
  
 +
Annealing Temp: 55
  
 
+
Cycles: 25
 
+
 
+
*H20 8.05μl
 
+
*10X Buffer 1.5μl
 
+
*MgCl2 (25mM) 1.2μl
 
+
*dNTPs (.25mM) 1.2μl
 
+
*356R 0.6μl
 
+
*356F 0.25μl
 
+
*Taq 0.2μl
AAT 300 - FAM
 
 
 
 
 
 
 
• Annealing Temp 55
 
• Cycles 22
 
 
 
 
 
 
 
    per sample x15 x110
 
• H20 5.05μl 75.75μl 555.5μl
 
• 10X Buffer 1.5μl 22.5μl 165μl
 
• MgCl2 (25mM) 1.2μl 18μl 132μl
 
• dNTPs (.25mM) 1.2μl 18μl 132μl
 
• 300R 0.6μl 9μl 66μl
 
• 300F 0.25μl 3.75μl 27.5μl
 
• Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AAT 308 - VIC
 
 
 
 
 
 
 
• Annealing Temp 55
 
• Cycles 24
 
 
 
 
 
 
 
    per sample x15 x110
 
• H20 5.05μl 75.75μl 555.5μl
 
• 10X Buffer 1.5μl 22.5μl 165μl
 
• MgCl2 (25mM) 1.2μl 18μl 132μl
 
• dNTPs (.25mM) 1.2μl 18μl 132μl
 
• 308R 0.6μl 9μl 66μl
 
• 308F 0.25μl 3.75μl 27.5μl
 
• Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AAT 312 - NED
 
 
 
 
 
 
 
• Annealing Temp 51
 
• Cycles 30
 
 
 
 
 
    per sample x15 x110
 
• H20 2.95μl 44.25μl 324.5μl
 
• 10X Buffer 1.5μl 22.5μl 165μl
 
• MgCl2 (25mM) 1.8μl 27μl 198μl
 
• KCl (0.5M) 1.5μl 22.5μl 165μl
 
• dNTPs (.25mM) 1.2μl 18μl 132μl
 
• 312R 0.6μl 9μl 66μl
 
• 312F 0.25μl 3.75μl 27.5μl
 
• Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AAT 333 - VIC
 
 
 
 
 
 
 
• Annealing Temp 51
 
• Cycles 30
 
 
 
 
 
 
 
    per sample x15 x110
 
• H20 3.85μl 57.75μl 423.5μl
 
• 10X Buffer 1.5μl 22.5μl 165μl
 
• MgCl2 (25mM) 2.4μl 36μl 264μl
 
• dNTPs (.25mM) 1.2μl 18μl 132μl
 
• 333R 0.6μl 9μl 66μl
 
• 333F 0.25μl 3.75μl 27.5μl
 
• Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AAT 350 - FAM
 
 
 
 
 
 
 
• Annealing Temp 52
 
• Cycles 28
 
 
 
 
 
    per sample x15 x110
 
• H20 5.05μl 75.75μl 555.5μl
 
• 10X Buffer 1.5μl 22.5μl 165μl
 
• MgCl2 (25mM) 1.2μl 18μl 132μl
 
• dNTPs (.25mM) 1.2μl 18μl 132μl
 
• 350R 0.6μl 9μl 66μl
 
• 350F 0.25μl 3.75μl 27.5μl
 
• Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AAT 356- VIC
 
 
 
 
 
• Annealing Temp 55
 
Cycles 25
 
 
 
 
 
 
 
 
 
    per sample x15 x110
 
H20 5.05μl 75.75μl 555.5μl
 
10X Buffer 1.5μl 22.5μl 165μl
 
MgCl2 (25mM) 1.2μl 18μl 132μl
 
dNTPs (.25mM) 1.2μl 18μl 132μl
 
356R 0.6μl 9μl 66μl
 
356F 0.25μl 3.75μl 27.5μl
 
Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
  
  

Latest revision as of 20:37, 12 December 2006

Below are the recipes that have been fine-tuned for Mimulus guttatus

AAT 9

Dye: VIC

Annealing Temp: 55

Cycles: 26

  • H20 8.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (2.5mM) 1.2μl
  • AAT9R 0.6μl
  • AAT9F 0.25μl
  • Taq 0.2μl

AAT 39

Dye: FAM

Annealing Temp 53

Cycles 25

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 39R 0.6μl
  • 39F 0.25μl
  • Taq 0.2μl


AAT 211

Dye: FAM

Annealing Temp: 53

Cycles: 22

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 211R 0.6μl
  • 211F 0.25μl
  • Taq 0.2μl


AAT 217

Dye: FAM

Annealing Temp: 54

Cycles: 27


  • H20 8.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 217R 0.6μl
  • 217F 0.25μl
  • Taq 0.2μl

AAT 222

Dye: NED

Annealing Temp 53

Cycles: 30

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 222R 0.6μl
  • 222F 0.25μl
  • Taq 0.2μl

AAT 225

Dye: VIC

Annealing Temp: 53

Cycles: 32

  • H20 8.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 225R 0.6μl
  • 225F 0.25μl
  • Taq 0.2μl

AAT 230

Dye: VIC

Annealing Temp: 54

Cycles: 28


  • H20 6.85μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 2.4μl
  • dNTPs (.25mM) 1.2μl
  • 230R 0.6μl
  • 230F 0.25μl
  • Taq 0.2μl

AAT 233

Dye: FAM

Annealing Temp: 53

Cycles: 28

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 233R 0.6μl
  • 233F 0.25μl
  • Taq 0.2μl

AAT 240

Dye: FAM

Annealing Temp: 54

Cycles: 25

  • H20 7.45μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • dNTPs (.25mM) 1.2μl
  • 240R 0.6μl
  • 240F 0.25μl
  • Taq 0.2μl

AAT 242

Dye: FAM

Annealing Temp: 49

Cycles: 32


  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 242R 0.6μl
  • 242F 0.25μl
  • Taq 0.2μl


AAT 261

Dye: VIC


Annealing Temp: 49

Cycles: 30


  • H20 4.45μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • dNTPs (.25mM) 1.2μl
  • 261R 0.6μl
  • 261F 0.25μl
  • Taq 0.2μl


AAT 265

Dye: NED


Annealing Temp: 55

Cycles: 20

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 265R 0.6μl
  • 265F 0.25μl
  • Taq 0.2μl


AAT 267

Dye: VIC


Annealing Temp: 49

Cycles: 30

  • H20 6.85μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 2.4μl
  • dNTPs (.25mM) 1.2μl
  • 267R 0.6μl
  • 267F 0.25μl
  • Taq 0.2μl

AAT 272

Dye: NED


Annealing Temp: 54

Cycles: 30

  • H20 4.45μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • dNTPs (.25mM) 1.2μl
  • 272R 0.6μl
  • 272F 0.25μl
  • Taq 0.2μl


AAT 278

Dye: NED

Annealing Temp: 53

Cycles: 24



  • H20 5.95μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • KCl (0.5M) 1.5μl
  • dNTPs (.25mM) 1.2μl
  • 278R 0.6μl
  • 278F 0.25μl
  • Taq 0.2μl

AAT 283

Dye: VIC

Annealing Temp: 52

Cycles: 29

  • H20 3.85μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 2.4μl
  • dNTPs (.25mM) 1.2μl
  • 283R 0.6μl
  • 283F 0.25μl
  • Taq 0.2μl


AAT 296

Dye: NED

Annealing Temp: 53

Cycles: 28


  • H20 2.1μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • KCl (.5M) 1.5μl
  • dNTPs (.25mM) 1.2μl
  • 296R 1.2μl
  • 296F 0.5μl
  • Taq 0.2μl


AAT 300

Dye: FAM

Annealing Temp 55

Cycles 22


  • H20 8.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 300R 0.6μl
  • 300F 0.25μl
  • Taq 0.2μl

AAT 308

Dye: VIC

Annealing Temp: 55

Cycles: 24

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 308R 0.6μl
  • 308F 0.25μl
  • Taq 0.2μl


AAT 312

Dye: NED

Annealing Temp: 51

Cycles: 30

  • H20 2.95μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • KCl (0.5M) 1.5μl
  • dNTPs (.25mM) 1.2μl
  • 312R 0.6μl
  • 312F 0.25μl
  • Taq 0.2μl


AAT 333

Dye: VIC

Annealing Temp: 51

Cycles: 30

  • H20 3.85μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 2.4μl
  • dNTPs (.25mM) 1.2μl
  • 333R 0.6μl
  • Taq 0.2μl


AAT 350

Dye: FAM

Annealing Temp: 52

Cycles: 28

  • H20 5.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 350R 0.6μl
  • 350F 0.25μl
  • Taq 0.2μl


AAT 356

Dye: VIC

Annealing Temp: 55

Cycles: 25

  • H20 8.05μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.2μl
  • dNTPs (.25mM) 1.2μl
  • 356R 0.6μl
  • 356F 0.25μl
  • Taq 0.2μl





AAT 364- NED


• Annealing Temp 53 • Cycles 30



    per sample	x15		x110	

• H20 3.7μl 55.5μl 407μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.8μl 27μl 198μl • KCl (.5M) 0.75μl 11.25μl 82.5μl • dNTPs (.25mM) 1.2μl 18μl 132μl • 364R 0.6μl 9μl 66μl • 364F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl













AAT 367 - VIC


• Annealing Temp 54 • Cycles 22


    per sample	x15		x110	

• H20 4.45μl 66.75μl 489.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.8μl 27μl 198μl • dNTPs (.25mM) 1.2μl 18μl 132μl • 367R 0.6μl 9μl 66μl • 367F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl











AAT 372- NED


• Annealing Temp 55 • Cycles 25



    per sample	x15		x110	

• H20 4.45μl 66.75μl 621.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 0.6μl 9μl 66μl • dNTPs (.25mM) 1.2μl 18μl 132μl • 372R 0.6μl 9μl 66μl • 372F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl










AAT 374 - FAM


• Annealing Temp 53 • Cycles 28


    per sample	x15		x110	

• H20 4.45μl 66.75μl 489.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.8μl 27μl 198μl • dNTPs (.25mM) 1.2μl 18μl 132μl • 374R 0.6μl 9μl 66μl • 374F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl











AG 19- NED


• Annealing Temp 53 • Cycles 30



    per sample	x15		x110	

• H20 5.65μl 84.75μl 621.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 0.6μl 9μl 66μl • dNTPs (.25mM) 1.2μl 18μl 132μl • AG19R 0.6μl 9μl 66μl • AG19F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl










ap3-VIC


• Annealing Temp 53 • Cycles 25


    per sample	x15		x110	

• H20 5.1μl 76.5μl 561μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.2μl 18μl 132μl • dNTPs (.25mM) 1.2μl 18μl 132μl • AP3R 0.6μl 9μl 66μl • AP3F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl










lfy- NED


• Annealing Temp 52 • Cycles 19


    per sample	x15		x110	

• H20 5.05μl 75.75μl 555.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.2μl 18μl 132μl • dNTPs (.25mM) 1.2μl 18μl 132μl • LFYR 0.6μl 9μl 66μl • LFYF 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl







    CYCB - FAM


• Annealing temp 62 • cycles 16


    per sample	x15		x110	

• H20 5.05μl 75.75μl 555.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.2μl 18μl 132μl • dNTPs (.25mM) 1.2μl 18μl 132μl • CYCBR 0.6μl 9μl 66μl • CYCBF 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl








    CYCA - FAM


• Annealing temp 58 • cycles 37


    per sample	x15		x110	

• H20 4.05μl 60.75μl 445.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.8μl 27μl 198μl • dNTPs (.25mM) 1.5μl 22.5μl 165μl • CYCAR 0.65μl 9.75μl 71.5μl • CYCAF 0.3μl 4.5μl 33μl • Taq 0.2μl 3μl 22μl