Mimulus:Microsatellite recipes

The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.

The original reference for the AAT markers: Kelly and Willis 1998
Check out the Mimulus Biological Resource Center for map location of these microsatellies.
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.
Contact David Lowry (dbl2@duke.edu) or other members of the Mimulus Community for more inforamtion about these markers.

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

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

H20 3.85μl
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 2.95μl
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 5.05μl
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 5.05μl
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

Mimulus:Microsatellite recipes

Contents

AAT 9

AAT 39

AAT 211

AAT 217

AAT 222

AAT 225

AAT 230

AAT 233

AAT 240

AAT 242

AAT 261

AAT 265

AAT 267

AAT 272

AAT 278

AAT 283

AAT 296

AAT 300

AAT 308

AAT 312

AAT 333

AAT 350

AAT 356

Navigation menu

Page actions

Page actions

Personal tools

Navigation

Search

research

Tools