7/17/23, 8:21 AM Primer3 Output (primer3_results.cgi release 4.1.

0)

Primer3 Output

PRIMER PICKING RESULTS FOR

Template masking not selected

No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any_th 3'_th hairpin seq
LEFT PRIMER 1580 20 58.99 55.00 0.00 0.00 0.00 CCACGGATGTCATTGCTCAG
RIGHT PRIMER 1806 20 58.97 50.00 0.00 0.00 0.00 TTTCACTTTCAGGGTTGCCG
SEQUENCE SIZE: 2082
INCLUDED REGION SIZE: 2082

PRODUCT SIZE: 227, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

1 ATGAAACGACATCTGAACACCAGCTACAGGCTGGTATGGAATCACATTACGGGCACCCTG

61 GTGGTGGCCTCCGAACTGGCCCGCTCACGGGGAAAACGCGCTGGTGTGGCGGTTGCGCTG

121 TCTCTTGCTGCTGTCACATCAGTCCCGGCACTGGCTGCTGACAAGGTTGTACAGGCGGGA

181 GAAACCGTGAACGACGGAACACTGACAAATCATGACAACCAGATTGTCCTCGGTACGGCC

241 AACGGAATGACCATCAGTACCGGTCTGGAGTATGGGCCGGATAACGAGGCCAATACCGGC

301 GGACAATGGATACAAAATGGCGGTATCGCCAACAACACTACTGTCACCGGTGGTGGTCTT

361 CAGAGAGTGAATGCCGGAGGAAGCGTTTCAGACACGGTTATCAGTGCCGGAGGCGGACAG

421 AGCCTTCAGGGGCAGGCAGTGAACACCACTCTGAACGGCGGTGAGCAGTGGGTACATGAA

481 GGCGGGATTGCAACGGGTACCGTCATTAATGAGAAGGGCTGGCAGGCCATCAAATCCGGC

541 GCAGTGGCAACCGACACGGTTGTGAATACCGGCGCGGAAGGGGGACCGGATGCGGAAAAT

601 GGTGATACCGGGCAGACCGTCTACGGAGATGCCGTACGCACCACCATCAATAAAAATGGT

661 CGTCAGATTGTGGCTGCTGAAGGAACGGCAAATACCACTGTGGTTTATGCCGGCGGCGAC

721 CAGACTGTACATGGTCACGCACTGGATACCACGCTGAATGGGGGGTACCAGTATGTGCAC

781 AACGGCGGTACGGCGTCTGGCACTGTTGTAAACAGTGACGGCTGGCAGATTGTCAAAAAC

841 GGGGGGGTGGCCGGGAATACCACCGTTAATCAGAAGGGCAGACTGCAGGTGGACGCCGGT

901 GGTACAGCCACGAATGTCACCCTGAAGCAGGGCGGCGCACTGGTTACCAGTACGGCTGCA

961 ACCGTTACCGGCATAAACCGCCTGGGAGCATTCTCTGTTGTGGAGGGTAAAGCTGATAAT

1021 GTCGTACTGGAAAATGGCGGACGCCTGGATGTGCTGACCGGACACACAGCCACTAATACC

1081 CGCGTGGATGATGGCGGAACGCTGGATGTCCGCAACGGTGGCACCGCCACCACCGTATCC

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1141 ATGGGAAATGGCGGTGTACTGCTGGCCGATTCCGGTGCCGCTGTCAGTGGTACCCGGAGC

1201 GACGGAAAGGCATTCAGTATCGGAGGCGGTCAGGCGGATGCCCTGATGCTGGAAAAAGGC

1261 AGTTCATTCACGCTGAACGCCGGTGATACGGCCACGGATACCACGGTAAATGGCGGACTG

1321 TTCACCGCCAGGGGCGGCACACTGGCGGGCACCACCACGCTGAATAACGGCGCCATACTT

1381 ACCCTTTCCGGGAAGACGGTGAACAACGATACCCTGACCATCCGTGAAGGCGATGCACTC

1441 CTGCAGGGAGGCTCTCTCACCGGTAACGGCAGCGTGGAAAAATCAGGAAGTGGCACACTC

1501 ACTGTCAGCAACACCACACTCACCCAGAAAGCCGTCAACCTGAATGAAGGCACGCTGACG

1561 CTGAACGACAGTACCGTCACCACGGATGTCATTGCTCAGCGCGGTACAGCCCTGAAGCTG
>>>>>>>>>>>>>>>>>>>>

1621 ACCGGCAGCACTGTGCTGAACGGTGCCATTGACCCCACGAATGTCACTCTCGCCTCTGGT

1681 GCCACCTGGAATATTCCCGATAACGCCACGGTGCAGTCGGTGGTGGATGACCTCAGCCAT

1741 GCCGGACAGATTCATTTCACCTCCACCCGCACAGGGAAGTTCGTACCGGCAACCCTGAAA
<<<<<<<<<<<<<<

1801 GTGAAAAACCTGAACGGACAGAATGGCACCATCAGCCTGCGTGTACGCCCGGATATGGCA
<<<<<<

1861 CAGAACAATGCTGACAGACTGGTCATTGACGGTGGCAGGGCAACCGGAAAAACCATCCTG

1921 AACCTGGTGAACGCCGGCAACAGTGCGTCGGGGCTGGCGACCAGCGGTAAGGGTATTCAG

1981 GTGGTGGAAGCCATTAACGGTGCCACCACGGAGGAAGGGGCCTTTATCCAGGGGAATAAG

2041 CTGCAGGCCGGTGCCTTTAACTACTCCCTCAACCGGGACAGT

KEYS (in order of precedence):

>>>>>> left primer
<<<<<< right primer

ADDITIONAL OLIGOS
start len tm gc% any_th 3'_th hairpin seq

1 LEFT PRIMER 587 20 58.98 50.00 0.00 0.00 0.00 CGGATGCGGAAAATGGTGAT

RIGHT PRIMER 747 20 59.10 50.00 0.00 0.00 0.00 ATCCAGTGCGTGACCATGTA
PRODUCT SIZE: 161, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

2 LEFT PRIMER 1646 20 58.91 55.00 0.00 0.00 0.00 CCATTGACCCCACGAATGTC

RIGHT PRIMER 1893 20 58.95 50.00 7.91 0.00 0.00 ACCGTCAATGACCAGTCTGT
PRODUCT SIZE: 248, PAIR ANY_TH COMPL: 14.36, PAIR 3'_TH COMPL: 0.00

3 LEFT PRIMER 725 20 58.93 55.00 0.00 0.00 0.00 CTGTACATGGTCACGCACTG

RIGHT PRIMER 921 20 58.92 55.00 0.00 0.00 0.00 GGTGACATTCGTGGCTGTAC
PRODUCT SIZE: 197, PAIR ANY_TH COMPL: 11.55, PAIR 3'_TH COMPL: 11.55

4 LEFT PRIMER 729 20 59.10 50.00 0.00 0.00 0.00 ACATGGTCACGCACTGGATA

RIGHT PRIMER 930 20 58.92 55.00 0.00 0.00 0.00 CTGCTTCAGGGTGACATTCG
PRODUCT SIZE: 202, PAIR ANY_TH COMPL: 16.08, PAIR 3'_TH COMPL: 2.57

Statistics
con too in in not no tm tm high high high high
sid many tar excl ok bad GC too too any_th 3'_th hair- poly end
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ered Ns get reg reg GC% clamp low high compl compl pin X stab ok
Left 11307 0 0 0 0 971 0 1555 5017 0 0 16 149 0 3599
Right 11306 0 0 0 0 882 0 1545 5087 0 1 17 149 0 3625
Pair Stats:
considered 1043, unacceptable product size 1034, primer in pair overlaps a primer in a better pair 82, ok 9
libprimer3 release 2.4.0

(primer3_results.cgi release 4.1.0)

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