Coding Sequence for Primer:
agt gggagatttt tcacggtcaa gctcccagtt
361 gctcttgatc ctgggtccaa gatctcggtc gttgtggaaa ctgtctacac ccatgtgctt
421 catccgtatc cgactcagat aactcagtca gagaaacagt tcgtcgtgtt tgagggcaac
481 cattatttct actctcccta tccaacaaag acccagacca tgcgagtgaa actggcgtcc
541 cgaaatgtgg aaagctatac caagctgggg aacccctcga gatctgagga cgtccttgat
601 tatgggcctt ttaaagacat ccctgcctac agtcagg

Primer 3 Output

WARNING: Numbers in input sequence were deleted.
PRIMER PICKING RESULTS FOR MGT M33

No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 24 20 59.96 55.00 4.00 4.00 gctcccagttgctcttgatc
RIGHT PRIMER 107 20 60.49 55.00 4.00 0.00 gtcggatacggatgaagcac
SEQUENCE SIZE: 310
INCLUDED REGION SIZE: 310

PRODUCT SIZE: 84, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 4.00

1 agtgggagatttttcacggtcaagctcccagttgctcttgatcctgggtccaagatctcg
>>>>>>>>>>>>>>>>>>>>

61 gtcgttgtggaaactgtctacacccatgtgcttcatccgtatccgactcagataactcag
<<<<<<<<<<<<<<<<<<<<

121 tcagagaaacagttcgtcgtgtttgagggcaaccattatttctactctccctatccaaca

181 aagacccagaccatgcgagtgaaactggcgtcccgaaatgtggaaagctataccaagctg

241 gggaacccctcgagatctgaggacgtccttgattatgggccttttaaagacatccctgcc

301 tacagtcagg

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

ADDITIONAL OLIGOS
start len tm gc% any 3' seq

1 LEFT PRIMER 26 20 59.80 50.00 4.00 0.00 tcccagttgctcttgatcct
RIGHT PRIMER 109 20 59.66 55.00 4.00 2.00 gagtcggatacggatgaagc
PRODUCT SIZE: 84, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 4.00

2 LEFT PRIMER 25 20 59.80 55.00 4.00 2.00 ctcccagttgctcttgatcc
RIGHT PRIMER 109 20 59.66 55.00 4.00 2.00 gagtcggatacggatgaagc
PRODUCT SIZE: 85, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 5.00

3 LEFT PRIMER 24 20 59.96 55.00 4.00 4.00 gctcccagttgctcttgatc
RIGHT PRIMER 108 20 60.62 50.00 4.00 1.00 agtcggatacggatgaagca
PRODUCT SIZE: 85, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 4.00

4 LEFT PRIMER 24 20 59.96 55.00 4.00 4.00 gctcccagttgctcttgatc
RIGHT PRIMER 98 20 59.37 50.00 4.00 2.00 ggatgaagcacatgggtgta
PRODUCT SIZE: 75, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 4.00

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 651 0 0 0 0 0 342 206 1 0 27 13 62
Right 614 0 0 0 0 0 369 149 2 0 42 10 42
Pair Stats:
considered 333, unacceptable product size 323, tm diff too large 2, ok 8
primer3 release 1.0

(primer3_www_results.cgi v 0.4)

Check Primers in Blast Search

LEFT PRIMER 24 20 59.96 55.00 4.00 4.00 gctcccagttgctcttgatc
RIGHT PRIMER 107 20 60.49 55.00 4.00 0.00 gtcggatacggatgaagcac

Primers OK to Test