7/17/23, 9:19 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 664 20 58.67 50.00 5.06 0.00 0.00 TCGGCATCTGGATCGATTCT
RIGHT PRIMER 912 20 59.53 50.00 0.00 0.00 0.00 TTCCGGCAGAAGGTTCAGTT
SEQUENCE SIZE: 1356
INCLUDED REGION SIZE: 1356

PRODUCT SIZE: 249, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

1 TTTATCCGTTTTGTATTGTAATAATCTATATATTTTTCTAATTCTATTTTGAAGTGCTCT

61 ACACTTTCAAACTCTTTTATGTAGAGAAACTCAGACTTCATGATTCCAAAGAAATTCTCC

121 ATTACTGCGTTGTCATAACAGTTGCCTTTACGAGACATACTTTGTACGATACCTCTTTTT

181 TGTAATGCGTGACGGTATTGTTTCATTTGATAATGCCACCCTTGATCTGAATGCATCATT

241 AGTTGGTGATTTTCAGGTAAACCTTCTAATGCTGTCTCTAACATTTCTGAAACAAGTGAA

301 TACGTCGGTCTAGAGCCAATTGTATAGGTAATGATTTCACCATTATACAAGTCTAATACA

361 GGTGATAAATAAAGTTTCTCACCGAATAATTTAAATTCTGTAATATCGGTTACCCATTTT

421 TCATTTGGCGTATCTGCCGTAAATTTACGCTCTAAAATATGAGGTGCAATTTTACCAACT

481 GTTCCTTTATAAGATTTATATTTTTTCATACGTACAAGACATTTTAAACCTAACTCTTTC

541 ATAATTCGTTGCACTTTTTTATGATTCACTATTTGATTACGATTGGCTAACTCATTACGA

601 ATACGACGATATCCATAACGACCTTCGTGTTCATCATAAATCGCTTTAATTTCTACTTTT

661 AAATCGGCATCTGGATCGATTCTATCCATTTGTTTTACATAAAAATAATAGGTGCTTCGA
>>>>>>>>>>>>>>>>>>>>

721 GGGATATCCGCTAACTTTACTAATTCTTTCACTTGAAACTCATGCCTTAATTCATAGACT

781 ACTTGTGCCTTGTCTTGTTTGGTGATTTTTCCTTGTTTTGAACTAAGGCATTCAACTTTT

841 TTAAATACGCATTTTCCATACGTAAGCGTTCTAATTCAGCTTGTAACGCTTCAACTGAAC
<<<<<<<<

901 CTTCTGCCGGAATCTTTTTAGTATCATGTTTTGTGTTTTTATTTTTCATGGATGGACGCC
<<<<<<<<<<<<

961 CCTTTTTCTTTGGTTCAAGGGCATCTAAGCCACCTACTTCAAATTGCTTCTTCCATTGAT

1021 AAACCGTTGTAAAAGAAGGAACCATGAATATAGCCGCTGTATCCATTAACGATATACCCG

1081 ATTGAGCCATAAAATTTAGTACGTCTAGTTTAAACTCTGCGGTGTAATTTGTATAGACCT

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7/17/23, 9:19 AM Primer3 Output (primer3_results.cgi release 4.1.0)
1141 CGTGTAAACCTTCTTCTCCATGGAGTTCGTATAAGCGTAACCAGTATTGAAATTGTGCCT

1201 TTGTAATATGCATCTGTTTTGCATATTCTGCTTGAGAAATTATTTTCGGATTAAATCCTT

1261 GAACGAGCGTTAACTTTAAATCCTTCGTAAATTTTGTCATACAAAAAACTGCACCTCCAA

1321 TTGTTAGACTGTGTCTAACAATTGGGGTGCAGTTCA

KEYS (in order of precedence):

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

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

1 LEFT PRIMER 710 20 58.66 55.00 13.87 13.87 0.00 AGGTGCTTCGAGGGATATCC

RIGHT PRIMER 909 20 58.41 50.00 0.00 0.00 0.00 CGGCAGAAGGTTCAGTTGAA
PRODUCT SIZE: 200, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

2 LEFT PRIMER 893 20 59.53 50.00 5.05 0.00 0.00 AACTGAACCTTCTGCCGGAA

RIGHT PRIMER 1089 21 58.76 47.62 0.00 0.00 0.00 TGGCTCAATCGGGTATATCGT
PRODUCT SIZE: 197, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

3 LEFT PRIMER 784 20 58.15 45.00 0.00 0.00 0.00 TGTGCCTTGTCTTGTTTGGT

RIGHT PRIMER 990 20 57.87 50.00 0.00 0.00 0.00 GCTTAGATGCCCTTGAACCA
PRODUCT SIZE: 207, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

4 LEFT PRIMER 138 20 57.10 45.00 0.00 0.00 0.00 ACAGTTGCCTTTACGAGACA

RIGHT PRIMER 321 20 58.82 50.00 0.00 0.00 0.00 AATTGGCTCTAGACCGACGT
PRODUCT SIZE: 184, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

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
ered Ns get reg reg GC% clamp low high compl compl pin X stab ok
Left 7120 0 0 0 0 2105 0 4062 43 0 0 12 115 0 783
Right 7065 0 0 0 0 1922 0 4128 43 0 0 29 131 0 812
Pair Stats:
considered 1955, unacceptable product size 1949, primer in pair overlaps a primer in a better pair 224, ok 6
libprimer3 release 2.4.0

(primer3_results.cgi release 4.1.0)

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