Microsattelite recipes

The original reference for the AAT markers: Kelly and Willis 1998

Check out the Mimulus Biological Resource Center for map location of these microsattelies. The primers are archived in genbank. Contact [dbl2@duke.edu David Lowry] for more inforamtion about the microsattelite markers.

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