Sequence used for Primer 3 (415-706):
gtttac
421 ccagggttga tgataacagc tggccttatt cattggattt taaatacatt gaacataaca
481 gttcacataa gagatgtgtg tgtattcctt gcaccaactt ttagcggcct tacatccata
541 tctacgttcc tgctaactag agaactgtgg aaccaaggag caggacttct agctgcttgc
601 ttcattgcta tcgtaccagg gtacatatct cggtcagtgg cgggatcctt tgataatgaa
661 ggcattgcca tttttgcgct tcagttcact tactacttat gggtaa

Primer 3 Report:
PRIMER PICKING RESULTS FOR GT66M01

No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 106 20 59.57 45.00 4.00 2.00 tttagcggccttacatccat
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
SEQUENCE SIZE: 292
INCLUDED REGION SIZE: 292

PRODUCT SIZE: 71, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00

1 gtttacccagggttgatgataacagctggccttattcattggattttaaatacattgaac

61 ataacagttcacataagagatgtgtgtgtattccttgcaccaacttttagcggccttaca
>>>>>>>>>>>>>>>

121 tccatatctacgttcctgctaactagagaactgtggaaccaaggagcaggacttctagct
>>>>> <<<<<<<<<<<<<<<<<<<<<

181 gcttgcttcattgctatcgtaccagggtacatatctcggtcagtggcgggatcctttgat

241 aatgaaggcattgccatttttgcgcttcagttcacttactacttatgggtaa

KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer

ADDITIONAL OLIGOS
start len tm gc% any 3' seq

1 LEFT PRIMER 109 21 59.97 47.62 4.00 2.00 agcggccttacatccatatct
RIGHT PRIMER 174 20 60.63 55.00 3.00 2.00 aagtcctgctccttggttcc
PRODUCT SIZE: 66, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00

2 LEFT PRIMER 155 20 60.63 55.00 3.00 1.00 ggaaccaaggagcaggactt
RIGHT PRIMER 228 19 60.23 57.89 4.00 2.00 cgccactgaccgagatatg
PRODUCT SIZE: 74, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00

3 LEFT PRIMER 106 20 59.57 45.00 4.00 2.00 tttagcggccttacatccat
RIGHT PRIMER 179 21 60.52 57.14 4.00 2.00 gctagaagtcctgctccttgg
PRODUCT SIZE: 74, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00

4 LEFT PRIMER 110 20 59.03 50.00 4.00 2.00 gcggccttacatccatatct
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 67, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

5 LEFT PRIMER 109 20 59.03 50.00 4.00 2.00 agcggccttacatccatatc
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 68, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00

6 LEFT PRIMER 109 21 59.97 47.62 4.00 2.00 agcggccttacatccatatct
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 68, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00

7 LEFT PRIMER 106 20 59.57 45.00 4.00 2.00 tttagcggccttacatccat
RIGHT PRIMER 173 19 59.24 57.89 2.00 2.00 agtcctgctccttggttcc
PRODUCT SIZE: 68, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

8 LEFT PRIMER 109 21 59.97 47.62 4.00 2.00 agcggccttacatccatatct
RIGHT PRIMER 182 21 60.30 57.14 5.00 2.00 gcagctagaagtcctgctcct
PRODUCT SIZE: 74, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00

9 LEFT PRIMER 156 21 59.87 52.38 4.00 4.00 gaaccaaggagcaggacttct
RIGHT PRIMER 228 19 60.23 57.89 4.00 2.00 cgccactgaccgagatatg
PRODUCT SIZE: 73, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00

10 LEFT PRIMER 106 21 59.59 42.86 4.00 2.00 tttagcggccttacatccata
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 71, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00

11 LEFT PRIMER 163 21 60.30 57.14 5.00 5.00 ggagcaggacttctagctgct
RIGHT PRIMER 228 19 60.23 57.89 4.00 2.00 cgccactgaccgagatatg
PRODUCT SIZE: 66, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00

12 LEFT PRIMER 109 21 59.97 47.62 4.00 2.00 agcggccttacatccatatct
RIGHT PRIMER 179 21 60.52 57.14 4.00 2.00 gctagaagtcctgctccttgg
PRODUCT SIZE: 71, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00

13 LEFT PRIMER 109 20 59.03 50.00 4.00 2.00 agcggccttacatccatatc
RIGHT PRIMER 173 19 59.24 57.89 2.00 2.00 agtcctgctccttggttcc
PRODUCT SIZE: 65, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

14 LEFT PRIMER 159 21 60.52 57.14 4.00 3.00 ccaaggagcaggacttctagc
RIGHT PRIMER 228 19 60.23 57.89 4.00 2.00 cgccactgaccgagatatg
PRODUCT SIZE: 70, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00

15 LEFT PRIMER 109 21 59.97 47.62 4.00 2.00 agcggccttacatccatatct
RIGHT PRIMER 173 19 59.24 57.89 2.00 2.00 agtcctgctccttggttcc
PRODUCT SIZE: 65, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

16 LEFT PRIMER 106 19 59.27 47.37 4.00 0.00 tttagcggccttacatcca
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 71, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00

17 LEFT PRIMER 106 21 59.59 42.86 4.00 2.00 tttagcggccttacatccata
RIGHT PRIMER 179 21 60.52 57.14 4.00 2.00 gctagaagtcctgctccttgg
PRODUCT SIZE: 74, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00

18 LEFT PRIMER 155 19 59.24 57.89 2.00 2.00 ggaaccaaggagcaggact
RIGHT PRIMER 228 19 60.23 57.89 4.00 2.00 cgccactgaccgagatatg
PRODUCT SIZE: 74, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00

19 LEFT PRIMER 110 21 59.07 47.62 4.00 2.00 gcggccttacatccatatcta
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 67, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

20 LEFT PRIMER 108 21 59.07 47.62 4.00 2.00 tagcggccttacatccatatc
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
PRODUCT SIZE: 69, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00

21 LEFT PRIMER 106 21 59.59 42.86 4.00 2.00 tttagcggccttacatccata
RIGHT PRIMER 173 19 59.24 57.89 2.00 2.00 agtcctgctccttggttcc
PRODUCT SIZE: 68, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

22 LEFT PRIMER 106 19 59.27 47.37 4.00 0.00 tttagcggccttacatcca
RIGHT PRIMER 173 19 59.24 57.89 2.00 2.00 agtcctgctccttggttcc
PRODUCT SIZE: 68, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00

23 LEFT PRIMER 108 21 59.07 47.62 4.00 2.00 tagcggccttacatccatatc
RIGHT PRIMER 173 19 59.24 57.89 2.00 2.00 agtcctgctccttggttcc
PRODUCT SIZE: 66, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00

Statistics
con too in in no tm tm high high high
sid many tar excl bad GC too too any 3' poly end
ered Ns get reg GC% clamp low high compl compl X stab ok
Left 624 0 0 0 20 0 456 69 0 0 31 9 39
Right 610 0 0 0 2 0 467 64 3 0 37 8 29
Pair Stats:
considered 1203, unacceptable product size 1163, tm diff too large 16, ok 24
primer3 release 1.0

Primer Pair to test: Ok in Blast searches

VALIDATED PRIMER SEQUENCES:

LEFT PRIMER 106 20 59.57 45.00 4.00 2.00 tttagcggccttacatccat
RIGHT PRIMER 176 21 59.87 52.38 4.00 0.00 agaagtcctgctccttggttc
SEQUENCE SIZE: 292
INCLUDED REGION SIZE: 292

PRODUCT SIZE: 71, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00