Apple_SSR_Genotype_Volk_2020

Project Overview
Project Name Apple_SSR_Genotype_Volk_2020
Description Apple genotype data obtained in 2020 from Gayle Volk, National Laboratory for Genetic Resources Preservation, USDA. About 1500 Malus x domestica and a few other species have been genotyped using nine SSR markers.
Comments Extract DNA from leaves, PCR amplify in Perkin Elmer 9600 thermocycler. Multiplexed primer sets are Primer Mix 1 (GD12-A labeled with 6-FAM(blue), GD142-A labeled with PET(red), CH01h01-A labeled with NED(yellow/black)), Primer Mix 2 (GD147-A labeled with PET(red), GD15-A labeled with VIC(green), CH01f02-A labeled with NED(yellow/black)), and Primer Mix 3 (GD162-A labeled with PET(red), GD96-A labeled with VIC(green), CH02d08-A labeled with 6-FAM(blue)). PCR amplified 5uL reaction mixtures containing 2 uL (2-10 ng) DNA, 2.5 uL mastermix (Qiagen multiplex PCR kit), 0.5 uL primer mix (PM1, PM2, or PM3, all primers 0.5 uM). PCR cycle: 95C for 15 min; followed by 40 cycles of 94C (1 min), 55C (1.5 min), 72C (1 min); and a final extension at 72C for 10 min. Hold at 4C for 15 min, hold at 12C. . PCR products were separated using an ABI 3730 capillary sequencer (Thermo Fisher Scientific, Waltham, MA), and the Liz 500 size standard (Thermo Fisher Scientific, Waltham, MA). Control samples were Golden Delicious (PI 590184), Rome Beauty Law (PI 588850), and Cox's Orange Pippin (PI 588853). Resulting chromatograms were analyzed to determine microsatellite allele sizes using a partially automated binning system software (Gene Mapper version 5, Thermo Fisher Scientific). Automated binning was set to a range of ± 0.4 to ensure that alleles differing by a single base pair were correctly scored.
Sub type SSR
Crop apple
Project type genotype
Stocks
This project contains 3,079 stocks:
NameType
PI 589911 PLclone
PI 589912 .04 PLclone
PI 589913 .12 PLclone
PI 589914 .07 PLclone
PI 589915 .06 PLclone
PI 589916 .06 PLclone
PI 589917 .06 PLclone
PI 589918 .07 PLclone
PI 589920 .08 PLclone
PI 589922 .07 PLclone
PI 589923 .07 PLclone
PI 589924 .05 PLclone
PI 589925 .06 PLclone
PI 589926 .05 PLclone
PI 589927 .08 PLclone
PI 589929 .06 PLclone
PI 589930 .11 PLclone
PI 589931 .07 PLclone
PI 589932 .14 PLclone
PI 589944 .04 PLclone
PI 589945 .05 PLclone
PI 589956 .14 PLclone
PI 589957 .05 PLclone
PI 589958 .12 PLclone
PI 589959 .05 PLclone
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Contact
NameDetails
G. Volk
First name:Gayle
Last name:Volk
Institution:USDA-ARS
Address:NATIONAL CENTER FOR GENETIC RESOURCES PRESERVATION, 1111 SOUTH MASON STREET, FORT COLLINS, CO, 80521-4500
Email:gayle.volk@ars.usda.gov
Phone:19704953205