Difference between revisions of "Mimulus:Microsatellite recipes"

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(AAT 225)
(AAT 230)
Line 119: Line 119:
  
  
 +
==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
  
AAT 233- FAM
+
*H20 4.45μl
 
+
*10X Buffer 1.5μl
 
+
*MgCl2 (25mM) 1.8μl
• Annealing Temp 53
+
*dNTPs (.25mM) 1.2μl
• Cycles 28
+
*240R 0.6μl
 
+
*240F 0.25μl
 
+
*Taq 0.2μl
 
 
    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
 
• 233R 0.6μl 9μl 66μl
 
• 233F 0.25μl 3.75μl 27.5μl
 
• Taq 0.2μl 3μl 22μl
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AAT 240 - FAM
 
 
 
 
 
 
 
• Annealing Temp 54
 
• Cycles 25
 
 
 
 
 
 
 
    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
 
240R 0.6μl 9μl 66μl
 
240F 0.25μl 3.75μl 27.5μl
 
Taq 0.2μl 3μl 22μl
 
 
 
  
  

Revision as of 20:49, 19 February 2006

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

AAT 9

Dye: VIC

Annealing Temp: 55

Cycles: 26

  • H20 5.05μl
  • 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 5.05μl
  • 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 5.05μl
  • 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 3.85μl
  • 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 4.45μl
  • 10X Buffer 1.5μl
  • MgCl2 (25mM) 1.8μl
  • dNTPs (.25mM) 1.2μl
  • 240R 0.6μl
  • 240F 0.25μl
  • Taq 0.2μl









AAT 242- FAM


• Annealing Temp 49 • Cycles 32


    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 • 242R 0.6μl 9μl 66μl • 242F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl









AAT 261 - VIC


• Annealing Temp 49 • Cycles 30


    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 • 261R 0.6μl 9μl 66μl • 261F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl












AAT 265- NED


• Annealing Temp 55 • Cycles 20


    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 • 265R 0.6μl 9μl 66μl • 265F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl









AAT 267- VIC


• Annealing Temp 49 • 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 • 267R 0.6μl 9μl 66μl • 267F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl











AAT 272- NED


• Annealing Temp 54 • Cycles 30


    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 • 272R 0.6μl 9μl 66μl • 272F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl










AAT 278 - NED


• Annealing Temp 53 • Cycles 24


    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 • 278R 0.6μl 9μl 66μl • 278F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl










AAT 283 - VIC


• Annealing Temp 52 • Cycles 29



    per sample	x15		x110	

• H20 3.85μl 96.25μl 423.5μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 2.4μl 36μl 264μl • dNTPs (.25mM) 1.2μl 18μl 132μl • 283R 0.6μl 9μl 66μl • 283F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl










AAT 296 - NED


• Annealing Temp 53 • Cycles 28


    per sample	x15		x110	

• H20 2.1μl 31.5μl 231μl • 10X Buffer 1.5μl 22.5μl 165μl • MgCl2 (25mM) 1.8μl 27μl 198μl • KCl (.5M) 1.5μl 22.5μl 165μl • dNTPs (.25mM) 1.2μl 18μl 132μl • 296R ** 1.2μl 18μl 132μl ** • 296F ** 0.5μl 7.5μl 55μl ** • Taq 0.2μl 3μl 22μl










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







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