**gpw93005**

GCAAAGGGGGCAAGCACAGGCCAAGGCATCAGGCAAAGGTTACAGTTTCGGTGCAACAAG

ACCCAGACCAGCAGAGCGGAATGATAGAGGTCATCGTGGAGCCCAAGACGGTGTCGTCGT

CAGAGCCTTTGGCAGCCGAAGACCAACTTTAGTCAGGATAAGTGTACTAGATGTTCCCCT

TAGAAATGGCCAATTGTTGGCGCCCTTCCCGCTCATTATTTGGGAAGAGGCCCAGGGCCT

CCCTCTATAAATAGGACTAGCCACCACAGAGTAGGGGCATCTTGGAGACGGATTAATCAA

AGAGAGAGAGAGAGAGAGAGAGGCGACTGAATTCACCCCCGCAGTTCATCGCACCAGCTC

AAGAACAGCACTCGCGAGGCTGTTCTCCCTTTGTATTGTTCATCATCAGCCCCTGAGGCA

ATCCACCACACCACACTGGAGTAAGGTATTACACCGCAATGGTGGCCTGAACCAGTATAA

ATCTCGTGTCCCTTGTGTGTTCTTCTTTCTAGCCTAGATCCTTTGTGAGACT

**Forward** : AGCCACCACAGAGTAGGGG

**Reverse** : GTGTCTTCGAGTGCTGTTCTTGAGCTG

**SSR motif** :(AG)11

**Annealing temperature** : 60^o^C

**Allele size** :

**Chinese Spring** -- 119 bp

**Opata** -- 119 bp

**Synthetic 6x** -- 119 bp

**Chromosome NT analysis :**

**Comments** : Very strong amplification with alot of non-specific
background. Impossible to map on nulli-tetra's.

**Pre-Screen Image :**

![](media/image1.png){width="1.7277777777777779in" height="3.6in"}

**gpw93018**

ATCTTTCCCACTAATGTTGACTGAACTTCCATTGCTGCAACAAAACCTTTGGGAACAATC

TCCAGCCGAAGTTCCCTGAGATCCTCGGCTATCTCCTTTGGCAATCCTCCACTTATGAGG

GTATTCGTCCTTGTTTTCTTTATGTTGTGTCCTATTTCTCCACAAAGCTTGTATGCACAA

GGTTTCTTCATATCCTTTCCATAAGGCCTCAACTTCTGGGACACAGGACGAGATCTTTGA

AGAGTCAACTTAAATTCTTCCCAGGTGGTTACTCTTTGCCACCCATTGATGGTTTGGTAC

ATACTCCACCAAGTAGCAGCAACTCTTAAAAAGCACTGAAGAGCATGCTGGGTTATATCC

TTTCTGGCTTTAGGGTTATCCTCCATGTTCTGAATCCATCGGTCCGTATCCAAATGACTC

ATCGGTCTAGAAAATTACAAGCTCATGTCCATTCATCCTTTATTGGGTCCATGGGTTTTG

GGGTGGGATAAGAGAGAGAGAGAGATAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGGGAT

GCACACAATACAAACAATGGTTTTTGAAAAGACAGATTTTATTGTGACTG

**Forward primer** : CAAGCTCATGTCCATTCATCC

**Reverse primer** : GTGTCTTTTGTTTGTATTGTGTGCATCCC

**SSR motif** : (AG)23i

**Annealing temperature** :60^o^C

**Allele size** :

**Chinese Spring --** 94 bp

**Opata --** 94 bp

**Synthetic 6x --** 94 bp

**Chromosome NT analysis**:

**Comments** : Monomorphic in the 7 varieties screened. Very strong
amplification - impossible to map on the nulli-tetra's.

**Pre-Screen Image :**

![](media/image2.png){width="2.1118055555555557in" height="3.4in"}

**gpw95001**

TGGTTGTAAAACTATTTTATTAAAGACAATTTCAATTGGGTTGTAAAACTATTTTAATTT

TCAGACAAATATTTGGACGTGCAAACTAGTTTTGATGAAAATATGGGCATTTTTGGCCCT

GACGGACAGGATGGGGGCAAACGGATGCGGCCGCGCGCTGGGCGCACGATCACCGCATTC

CAGGACACGTCCGGACACGACCCCATTGCCCTATCCAAACGGATAGAATCCGGACAAAAC

GGACTTCCGTTTAGGGTCGCGCGGTGGTGGGGTTGGCCTTACGACGGCAGCGATGTGAGT

GGTGAGAGCATTCGGTTTGGCACCGATGGTGGGAGCGAGACATGGTCCAGGTCGTCCACG

CCCAGTGATTTTGTCCCTACGACGGGTGGCTTTCCGTCCGACGTGCGGTGTAATCCATCC

ATCGCTTTCCGCTTGCTCAAGAGTCCGGTAAGGCCGCCGTGCAGCCTCGCTCCTTCTTCT

CCCCTCCAACCTTTTCCCCCTGTTCTGCAAGCCCTTTCCGTCTCCTTCTGGCTGCTTCCT

CGCGCAGGTGCTATTTCTGAGCTTCCGAGGCGACAGAGCTGCACAAGAGCAGAGCAGAGC

ATAGGGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGATGAGGGAGA

TCATAAGCATCCACATCGGGCAGGCGGGCATCCAGGTGGGGAATTCCTGCTGGGAGCTCT

ACTGC

**Forward primer** : TTCTTCTCCCCTCCAACCTT

**Reverse primer** : GTGTCTTTGTGGATGCTTATGATCTCCC

**SSR motif** : (GA)23

**Annealing temperature** :50^o^C

**Allele size** :

**Chinese Spring** -- 182 and 192 bp

**Opata** -- 182 and 188 bp

**Synthetic 6x** -- 182 and 188 bp

**Chromosome NT analysis** : Upper locus of 192 bp, in Chinese Spring
maps to 4DL and the lower locus of 182 bp maps to either 2AL or 4DS.

**Comments** : Two loci, lower locus of 182 bp was monomorphic in the 7
varieties screened.

# Pre-Screen Image :

![](media/image3.png){width="1.3972222222222221in"
height="3.4555555555555557in"}

**gpw95005**

AAGTGGTTGTTAATAAGACCTAATCAACCTTATTAATGGATCATTTTTAGTGGTATGAAG

TTGATGAATTTAGTAAGCACTACCTTCTGTCCCTACCTTTTGTTTTCTGTTTTGTTAGTT

AAATAAAATAAAATGCCATGTTTTGTCTGTTTTCTGAATTTCCCGTGCAATAAGAAATGG

CCCAAAAATAAAAGTTCTCAGAATGCTCTGAAAATTTAGTACGATTTTTTTTGAATATTT

GAGAACTTCTGGTACAAATAATATCAGAGGGAGGTGCACTAGGTGAGCACAACCCACCTG

GGCGTGCCAGGGCCCCCAGGCGCGCCCTGGTGGGTTGTGGTCCCCACGTGGTCCCCTCAC

TTATCCCTTCACACCACATCATCACTTACCTCCACAAAAAATTCTCCATTGCTCTCTCTC

TCTCTCTCTCTCTCTCTCTCTCTCTCGTGTTCTTGCTCTCAAACCCGCGAATT

**Forward primer** : ACTAGGTGAGCACAACCCAC

**Reverse primer** : GTGTCTTCGGGTTTGAGAGCAAGAACA

**SSR motif** : (TC)17

**Annealing temperature** : 50^o^C

**Allele size** : **2A 3B 6B 7B Unlinked**

**Chinese Spring - -** 107 155 163

**Opata** 157 204 105 155 163

**Synthetic 6x** 158 - 101 153 -

**Chromosome ITMI map** : 2A (interval Xbcd152b-Xbdc543), 3B (interval
Xabg471b-Xbcd1095), 6B (Xfbb327-Xfbb59c), 7B (interval Xbcd310-Xksud27a)
and unlinked

**Chromosome NT analysis** : 144bp on 5AL, 140bp on 3AL, 136bp on 3D
and/or 5B, 130bp on 6AS, 115bp on 6AS and 89bp on 2DL.

**Comments** : Complex banding pattern.

**Pre-Screen Image :**

![](media/image4.png){width="1.5145833333333334in"
height="3.5194444444444444in"}

**gpw95010**

gcactcaacctatttataagttgggatccattccgcaaatgatgatagaatcatgtagaa

agagagagagagagagagagagagagagagagagagggagggagggagagagagagagag

atcgcctgcgtacctcctggagtgtgaagttggtgatgagatgagaac

**Forward primer** : CCATTCCGCAAATGATGATA

**Reverse primer** : GTGTCTTCACCAACTTCACACTCCAGG

**SSR motif** : (AG)17(AGGG)3(AG)7

**Annealing temperature** : 50^o^C

**Allele size** : **6A 6B 6D**

**Chinese Spring** 105 129 125

**Opata** 92+103 - 127

**Synthetic 6x** 107 137 134

**Chromosome ITMI map** : 6A (interval Xfba111b-Xmwg2053), 6B (interval
Xcdo507-Xfba42c) and 6D (interval Xbcd1510-Xksud27a)

**Comments** : Using an annealing temperature of 60oC only the locus on
chromosome 6D is amplified.

Chromosome 6A -- Opata has double-banded allele of 92+103 bp and
Synthetic 6x has a single band of 107 bp

Chromosome 6B - dominant marker with allele 137 amplified from Synthetic
6x

Chromosome 6D -- this is the main locus with alleles of 127 bp in Opata
and 134 bp in Synthetic 6x

**Pre-Screen Image :**

![](media/image5.png){width="1.4180555555555556in"
height="3.3354166666666667in"}

**gpw95016**

caccaacaagacatcaacaagacagccacaccgcggccatgcctggccactattgctgca

taggcatacactattcatccatcaccatcaacagtgtgcagtaccatcaatagtgtaact

tgtcccctgggcactagtatataaagtgacccagggggcaagccattgggccatggccac

aaggcctcacacaggctggcaagctaactagctatccatccatccaggatgaacacactc

cacgcatacatgctagctagcaagctagctagcacacacacacatccattcatccatcca

cccacgtccaagcacacacttgtgctcatatgcatatacgaggctagatgcctatggcat

tgacctcagatacagctccaggagcaaccctgtaccagatatacataccagatatactca

gacatgcaggagtaggggtattatctctccggagagccctgaacctgggtaaactagcgt

gtgctactcgcatacccactcccgggcctatcagcagcagtcttaccctcacactaagcc

cttgtggcatcag

**Forward primer** : CAGGATGAACACACTCCACG

**Reverse primer** : GTGTCTTGTGTGAGGGTAAGACTGCTGC

**SSR motif** : (CA)6

**Annealing temperature** :50^o^C

**Allele size** :

**Chinese Spring -** 312, 301, 277, 268, and 200 bp

**Opata** - 312, 304, 301, 277, 268, and 200 bp

**Synthetic 6x -** 312, 301, 277, 268, and 200 bp

**Comments** : Monomorphic, but couldn't assign bands to chromosomes
using the nulli-tetra's. Dominant marker allele of 304 bp wasn't mapped
in the ITMI population.

**Pre-Screen Image :**

![](media/image6.png){width="1.2902777777777779in"
height="4.423611111111111in"}

**gpw95022**

cttcaaatgctgaggatgacgacgagaaccaccaaaattattgagacactccggaacaat

taagaatagaaaggttgagtgaaaaaaatgaaaagaattttgaaagtgatcttggaataa

gcatatgactgatgaaattcattcttacgtcaaacttggaaataattagagaataactgc

agaaaaaattagaagagcaggtaagatcctggggataaacctgtcggttagggcccactg

aaaagaagcaccgttgaaaatgattcaagaagagagatgatacaccagagaaattgaaag

acttgaatgaggtgacaaccttgaattaattagaacacaaacacgaatcttctgagatgt

ccttagcactccgaaatgatgaaatggcgagaggtgaacgagcaagaggatacatttgag

ccgagggaaagaatatgatcaacactggatacttgaattgaatccactggagaagagaac

gagagagagagagagagagagagagagagagagagagagaatgacgaactagaatccacc

ggttgaaacaattaaggaaagaatgaagaggcttcacacgaacaaaatgatac

**Forward primer** : GATAAACCTGTCGGTTAGGGC

**Reverse primer** : GTGTCTTCCGGTGGATTCTAGTTCGTC

**SSR motif** : (GA)20

**Annealing temperature** : 60^o^C

**Allele size** :

**Chinese Spring -** 312 and 330 bp

**Opata -** 310 and 336 bp

**Synthetic 6x -** 310 and 332 bp

**Chromosome ITMI map** : 7D (interval Xbcd707-Xcdo775)

**Comments** : Two loci -- lower locus not polymorphic in ITMI, but
wasn't screened against the nulli-tetra's.

**Pre-Screen Image :**

![](media/image7.png){width="1.663888888888889in"
height="2.9194444444444443in"}

**gpw95024**

CAATTCAGTCCACACGCCCGACACAGATTCAGAAACAAGTTACGGCCGAACGCCACACAC

ACACACACACACACACACAGGCTCATTCAGATCCTATGGTTTCGCTTGACGTTGGCTGGG

CTGGTGGCTGCGGACATGGAACAACTCTTGGCTGTTACAGAGAGTTCATTTCTGAATATC

TCCTAGGTTTCATGACGCTGCCCGCTCTGATGTCTGCTTCTCCTTGTCTTCCTCTCCCAA

GAGTTTCCTCCGTTGCTGCATAAAGAACAAGAATTTGAATCTGATAGTGTAGTGTCAGTG

TTGGAGTAGATTCTGGTGGCTCATTGAAGTAGTGAACTTCAGTAGACATGCAAGATATCT

CTAGAACTTCAACTTCTGCTATTAAAACATGTGAGAACCCAACCTCCGTCCGAAATTACT

ACCCGTTTGAGCGACAAGTAATTTCGGATGGAGTACTACATGACCATCAGAAATTCAGAA

CCAAACCAATCTTGATCTTAGCTAACTAGTGGATTTGACTGTCATCCAGTTGCTGATTAC

CTTGTAATGCAGGAAGCCAATCAGCGAATGTGAATTCGGGGAACGAGCTGAGGTCGCGTA

CGGCTACTCGGAAAACGTCGGTGAGCACGGTCTTGGACTGCAGCATCACGGCCTTGAGGA

TCTCTTTGCCCTGCAGTTGCACCATCAGATCAAACACGTGACCTGCATCGCTCAGTTCCA

AGATGAAAATGGAAACGGAACAGTAGTAATTAAGCCACCTCCTTCAACCCAAGCTGCACG

**Forward primer** : AATTCAGTCCACACGCCC

**Reverse primer** : GTGTCTTAGCAGACATCAGAGCGGG

**SSR motif** : (CA)13

**Annealing temperature** : 60^o^C

**Allele size** :

**Chinese Spring** -- 250 bp

**Opata** -- 252 bp

**Synthetic 6x** -- 242 bp

**Chromosome ITMI map** : 3D (interval Xcdo407-Xbcd515)

**Comments** : Single locus

**ITMI population :**

![](media/image8.png){width="5.375694444444444in"
height="2.1034722222222224in"}

**gpw95027**

AAAAGTCTACAAACGATCCCCTATACTTGTGGGTTATCAGTATTCAATCCGATAAACGAA

ATCTCAAAGGGAAAACTCAATTCATCACAACAAGATAGAAATGGAGGAACACCATATGAT

CCAACTATATTGACAAAGCCCGCGATACATCAGGATCGTGACATCTCAAGAACACGAGAG

AGAGAGAGAGAGAGAGAGAGAGAGAGATTAAACACATAGCTACTGGTACAAACCCTCAGC

CCCGAGGGTGGACTACTCCCTCCTCATCATGGCGGTCAACGGGATGATGAAGATGGCCAC

CGGTGATGATTTCCCACTCCGACAGAGTACCAGAATGGGGTCTAGATTGGTTTTTCATGG

CTACAGAGGCTTGCGGCGGCGGAACTTCTGATTTAGGGTCATTTCTGATGGTTTTCCTAT

TTATAGGATTTTTTGGCG

**Forward primer** : TGACAAAGCCCGCGATAC

**Reverse primer** : GTGTCTTGCCATGATGAGGAGGGAGTA

**SSR motif** : (GA)16

**Annealing temperature** : 60^o^C

**Allele size** :

**Chinese Spring**

**Opata**

**Synthetic 6x**

**Chromosome NT analysis** :

**Comments** : Single, monomorphic locus with lots of stutter. Very
strong amplification - impossible to map on the nulli-tetra's.

**Pre-Screen Image :**

![](media/image9.png){width="1.6958333333333333in"
height="3.4319444444444445in"}

**gpw95032**

acgcgcgcgatgcgactggccgaacgtttcggccggtgcgctggcgtaaaccgtttacct

tataattagcgggtctcaccccaatgaaatttagagggaggggtgtgtgtgtgtgtgtta

aaacttatataggttaatcgtacatttaggattagtgaaaggggaggggggtaaattaga

ggatggacacatacgtgcgcaaggtgcagcctggataggtgctgcatgtaagtttactat

ttccgatactataaacgcgtgcagaccgcaactcatgcatcgttctcaaacaataaatct

ccaatacacaagaaggtttctgactcagctagcgaaatggctttcgcgctcaatgcatct

tgctacccttcgtctttctcgtcgctgctcccgtggcggatggcagcagcggtgatgata

ccaaggcggaggaatgttctgcctgtcattaaggcggttgcggtggctccgcccgccccg

gcgccggcgaagccagcagccgtgaggagccggcccgtgtcggtcaccatggccaagctc

atggctaagggcaaggtgcgtgcttatgtgtgtgcatgtgttgtacattgcatgtaacgg

acgtgtggactctgaccggatattgggg

**Forward primer** : GGGTCTCACCCCAATGAAAT

**Reverse primer** : GTGTCTTCATGCAGCACCTATCCAGG

**SSR motif** : (GT)8

**Annealing temperature** : 60^o^C

**Allele size** :

**Chinese Spring** -- 163 bp

**Opata** -- 163 bp

**Synthetic 6x** -- 165 bp

**Chromosome NT analysis** : 4D (interval Xksuf8b-Xfbb178b)

**Comments** : Single locus

**Pre-Screen Image :**

![](media/image10.png){width="1.663888888888889in" height="2.94375in"}