Primer3 Output (primer3_results.cgi release 4.1.0) https://primer3.ut.ee/cgi-bin/primer3/primer3web_results.

cgi

Primer3 Output

PRIMER PICKING RESULTS FOR DACHS1

Template masking not selected

Using mispriming library humrep_and_simple.txt
Using 1-based sequence positions
OLIGO start len tm gc% any_th 3'_th hairpin rep seq
LEFT PRIMER 221 20 58.93 50.00 0.00 0.00 0.00 11.00 TCTGGAGAGTTTGGCACCAT
RIGHT PRIMER 688 20 60.61 55.00 0.00 0.00 0.00 12.00 GGTGGGTGCATGGTCATTGA
SEQUENCE SIZE: 840
INCLUDED REGION SIZE: 840

PRODUCT SIZE: 468, PAIR ANY_TH COMPL: 13.51, PAIR 3'_TH COMPL: 0.00
TARGETS (start len
EXCLUDED REGIONS (start len

1 CCACACACAAATGATACCAGCTGCTTTATAGTATCTTCTTTTACTTTCTGTTTCTGTTTT

61 TCTTCCTTTCTCTCATAAGGTTCACTTGCTTTTGTGCACATGCTCACTCTCTCCTGTTGT

121 CTGTAAATCACCCCCATCCCCACGACACATGTATATTTATATGTGCATGCCCTTCTCTGC

181 AGATGGGGACCCATCAGGAGCCTTTGTCCTAGACCTTGCTTCTGGAGAGTTTGGCACCAT
>>>>>>>>>>>>>>>>>>>>

241 GCGGCCACTAGACAGAGAAGTGGAGCCAGCTTTCCAGCTGAGGATAGAGGCCCGGGATGG
XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

301 AGGCCAGCCAGCTCTCAGTGCCACGCTGCTTTTGACAGTGACAGTGCTGGATGCCAATGA
XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

361 CCATGCTCCAGCCTTTCCTGTGCCTGCCTACTCGGTGGAGGTGCCGGAGGATGTGCCTGC
XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

421 AGGGACCCTGCTGCTGCAGCTACAGGCTCATGACCCTGATGCTGGAGCTAATGGCCATGT
************************************************************

481 GACCTACTACCTGGGCGCCGGTACAGCAGGAGCCTTCCTGCTGGAGCCCAGCTCTGGAGA
************************************************************

541 ACTGCGCACAGCTGCAGCCTTGGACAGAGAACAGTGTCCCAGCTACACCTTTTCTGTGAG
XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

601 TGCAGTGGATGGTGCAGCTGCTGGGCCCCTAAGCACCACAGTGTCTGTCACCATCACGGT
XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

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Primer3 Output (primer3_results.cgi release 4.1.0) https://primer3.ut.ee/cgi-bin/primer3/primer3web_results.cgi

661 GCGCGATGTCAATGACCATGCACCCACCTTCCCCACCAGTCCTCTGCGCCTACGTCTGCC
<<<<<<<<<<<<<<<<<<<<

721 CCGCCCAGGCCCCAGCTTCAGTACCCCAACCCTGGCTCTGGCCACACTGAGAGCTGAAGA

781 TCGTGATGCTGGTGCCAATGCTTCCATTCTGTACCGGCTGGCAGGCACACCACCTCCTGG

KEYS (in order of precedence):

XXXXXX excluded region
****** target
>>>>>> left primer
<<<<<< right primer

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

1 LEFT PRIMER 218 20 59.12 55.00 0.00 0.00 0.00 12.00 GCTTCTGGAGAGTTTGGCAC
RIGHT PRIMER 691 19 59.93 57.89 0.00 0.00 0.00 11.00 GAAGGTGGGTGCATGGTCA
PRODUCT SIZE: 474, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

2 LEFT PRIMER 211 20 57.98 50.00 0.00 0.00 0.00 11.00 AGACCTTGCTTCTGGAGAGT
RIGHT PRIMER 705 19 60.23 63.16 0.00 0.00 0.00 11.00 AGAGGACTGGTGGGGAAGG
PRODUCT SIZE: 495, PAIR ANY_TH COMPL: 0.00, PAIR 3'_TH COMPL: 0.00

3 LEFT PRIMER 206 21 58.82 52.38 0.00 0.00 0.00 10.00 GTCCTAGACCTTGCTTCTGGA
RIGHT PRIMER 701 18 59.24 66.67 0.00 0.00 0.00 12.00 GACTGGTGGGGAAGGTGG
PRODUCT SIZE: 496, PAIR ANY_TH COMPL: 1.55, PAIR 3'_TH COMPL: 0.00

4 LEFT PRIMER 200 20 58.55 55.00 0.00 0.00 0.00 10.00 GCCTTTGTCCTAGACCTTGC
RIGHT PRIMER 693 18 60.36 66.67 0.00 0.00 0.00 12.00 GGGAAGGTGGGTGCATGG
PRODUCT SIZE: 494, 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 high
sid many tar excl ok bad GC too too any_th 3'_th hair- lib poly end
ered Ns get reg reg GC% clamp low high compl compl pin sim X stab ok
Left 1363 0 0 180 0 191 0 505 128 0 0 1 0 36 0 322
Right 1083 0 0 120 0 170 0 82 428 0 0 20 0 0 0 257
Pair Stats:
considered 322561, unacceptable product size 322553, high mispriming library similarity 2, primer in pair overlaps a primer in a better pair 5401, ok 6
libprimer3 release 2.4.0

(primer3_results.cgi release 4.1.0)

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