Mimulus:Microsatellite recipes

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(AAT 39)
Line 29: Line 29:
Cycles 25
Cycles 25
-
 
*H20 5.05μl
*H20 5.05μl

Revision as of 12:18, 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 - FAM


• Annealing Temp 53 • 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 • 211R 0.6μl 9μl 66μl • 211F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl












AAT 217 - FAM


• Annealing Temp 54 • Cycles 27


    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 • 217R 0.6μl 9μl 66μl • 217F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl











AAT 222- NED


• Annealing Temp 53 • Cycles 30


    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 • 222R 0.6μl 9μl 66μl • 222F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl






AAT 225- VIC


• Annealing Temp 53 • 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 • 225R 0.6μl 9μl 66μl • 225F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl












AAT 230 - VIC


• Annealing Temp 54 • Cycles 28


    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 • 230R 0.6μl 9μl 66μl • 230F 0.25μl 3.75μl 27.5μl • Taq 0.2μl 3μl 22μl






AAT 233- FAM


• Annealing Temp 53 • 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 • 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









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

Personal tools