7/17/23, 9:22 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
WARNING: Unrecognized base in input sequence

OLIGO start len tm gc% any_th 3'_th hairpin seq

LEFT PRIMER 1275 20 58.66 55.00 6.13 7.88 0.00 GCTAGTAATCGCGGATCAGC
RIGHT PRIMER 1507 20 58.07 50.00 9.78 0.00 0.00 TTATGAAGTTAGCGGCGGAC
SEQUENCE SIZE: 1552
INCLUDED REGION SIZE: 1552

PRODUCT SIZE: 233, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

1 AGGSYTSGGAAWTWKWWMMSSCGGGCTWTWGGSTTYCCGGATTTTTWAGGRATGGWCCCG

61 GGTTCGCCTTYAGCTTAGTKGGTTAAGGTAACGGCTTCACCCWAGGCCAACGATTSCGWA

121 CYSGAAACSKTKWKAKGGTGATTMGTKCCCACACTYGGGRTTSWGMCACGSYCYYWGTAA

181 CGTCCATTGTAGCRSGWRGGYAGCCASYAGGKMATTWWKSGGCMATGGAYGAAAATTCTG

241 ACGGAKCAWCSCCCRCSTTCCTTCCGGWKTGWTSAMGGGCAGTCACYTTMGRGTCGYMMA

301 ACTAAMTGWTGKYARSKAAGAWCAAGKGYTGMGYTSRWTAAGCTGGSACCTTRACSSWAC

361 ATCTMACSASACRAGCTSACGRCTAACYAYGYRCCASCWGYCRCKSTRMTCMCGWAGGWG

421 GCAAGCSYTATCTCYRGRRTTGTCAKWGGRYGTMAAGMSCKSGYAGGWGGTTTCTTCRMG

481 TCTGMTKYGAATTARMCCACAKGCTCMACCGCTTGTGMGGGYCMYYGKMAAYTGGGASMY

541 TTGAGTTKCAGCCWWGMGGAMMGTGGAMTCYCCAKGYGKAGYGSTGWAATGCGTWRASWT

601 MWGSASKAACACCAGTGGCGAAGGMRACTTYCTGGTCTRWMACTKAGCACTSAGKCGYKW

661 AMGCGYGKGGAGCWAMCAGGAKTAKMTAAYCCTGKTWGCTCCCCACGCCKTWMRCGCMTS

721 AGTGCTMAGTTACAGAYYAGARRGTCGCYTYCGCCACTGGTGTTMSTSCWKAWSTYWACG

781 CATTTMASCRCTMCRCMTGGGRAKTCCACKKYCGCWMKKCTGAMACTCAAAGTCTCCSAR

841 TTKMCRRKGRCCCKCACAAGCGGTKGAGCAYGTGGKYTAATTCRMAKCAGACGYKAAGAA

901 ACCACYTRCSMGSKCTTKACRYCCWMTGACAWYCCTAGAGATARSGCTTGCCWCCTWCGG

961 KAKYASMGYGRCWGSTGGTGCATSGTWGTCGTCAGCTCGTGKCKTGAGATSTTGGGTTAR

1021 GTMCCGTCAASGWGCGCAACSCTTGATCTYAGTTRCYAKCAYTTAGTTSKKCACTCTAAG

1081 GYRACWGMSKKTKACARACCSGAGGAAGGTGGGGATGMCKTCAAATCACTCAYGCSSCKT

1141 ATGACCTSSGCTACASACKTKCGTMCAWTGCGGAMGRTWCAMMKAGCTGCAWGACCKCSM
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7/17/23, 9:22 AM Primer3 Output (primer3_results.cgi release 4.1.0)

1201 GKWGGAGCTAATCTCATARRRCCSTGTCTCAGTTCSSAKTGTAGGCYGCAWCWCGCCTAC

1261 ATGMAGSTSGAATCGCTAGTAATCGCGGATCAGCWTGCCKYGGTGAAKMCGTTMCCKSRC
>>>>>>>>>>>>>>>>>>>>

1321 CAACTWGCTAAYRCRMCGCSSGTCACATCCAYRAGAGTTTGTAACASCCGAAGYCGGTGG

1381 SSTWWCMWTTTSGARCCAKSCGCGTTCAAARTGATTATCCGGTATTAGCCCCGGTTTCCC

1441 GGAGTTATCCCAGTCTTATGGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAAC
<<<<<<<<<<<<<

1501 TTCATAAGAGCAAGCTCTTAATCCATCGCTCGACTGCATTATAGCTGCCGCA
<<<<<<<

KEYS (in order of precedence):

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

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

1 LEFT PRIMER 1421 20 60.18 60.00 0.00 0.00 0.00 GGTATTAGCCCCGGTTTCCC

RIGHT PRIMER 1537 20 58.15 50.00 0.00 0.00 0.00 GCAGTCGAGCGATGGATTAA
PRODUCT SIZE: 117, 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 1713 1413 0 0 0 2 0 98 62 0 0 106 0 0 32
Right 2105 1331 0 0 0 6 0 268 192 0 0 23 0 0 285
Pair Stats:
considered 52142, unacceptable product size 52140, primer in pair overlaps a primer in a better pair 41192, ok 2
libprimer3 release 2.4.0

(primer3_results.cgi release 4.1.0)

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