Parameters 
Targets File: 
Non Targets File: 
Primers File: mergedPrimersN6.ml=20.Ml=25.txt
Primers Output File: 
Min Primer Length: 20
Max Primer Length: 25
Min Product Length: 75
Max Product Length: 200
Forward Primer: NONE
Reverse Primer: NONE
Range of positions for forward primers: 0 - 100000
Range of positions for reverse primers: 0 - 100000
Targets Mask: 11
Non Targets Mask: NONE
Degeneracy Mask: 1
Number of Source Sequences: 1
Min Coverage Targets: 100.00
Max Coverage Non Targets: 0.00
Max Self Score: 800.00
Max End Score: 300.00
Min GC Content: 25.00
Max GC Content: 75.00
GC Clamp: 0
Max Poly-X: 5
Primer Concentration: 0.000000800
Template Concentration: 0.000000000
Salt Concentration: 0.05000
Salt Correction Method: 1
Targets Min Melting Temp : 40.00
Targets Max Melting Temp: 100.00
Non Targets Max Melting Temp: 40.00
Non Targets Delta Temp: 0.00
Max Pair Temperature Difference: 40.00
Primers Label: N6
Dependent stats
Sequence for ranking: NONE
Targets: 0
Non Targets: 0
Load time: 0
Running process for forward primers
Build tables time: 0
Forward_N6_S1_3 AGATAATATGCATTTCAGCTACAGG 58 42.13 49.98 56.96 33.19 100.00% 0.00%
Forward_N6_S1_4 GATAATATGCATTTCAGCTACAGGA 63 41.32 48.97 55.75 29.80 100.00% 0.00%
Forward_N6_S1_7 TGCATTTCAGCTACAGGAATGACAC 66 45.10 53.60 59.88 28.27 100.00% 0.00%
Forward_N6_S1_8 GCATTTCAGCTACAGGAATGACACT 67 46.55 54.67 60.74 26.81 100.00% 0.00%
Forward_N6_S1_9 CATTTCAGCTACAGGAATGACACTA 68 42.57 52.31 57.96 25.68 100.00% 0.00%
Forward_N6_S1_10 ATTTCAGCTACAGGAATGACACTAT 69 41.59 50.96 57.77 22.78 100.00% 0.00%
Forward_N6_S1_11 TTTCAGCTACAGGAATGACACTATC 70 41.17 50.31 56.32 25.17 100.00% 0.00%
Forward_N6_S1_12 CAGCTACAGGAATGACACTATCGGT 73 46.04 53.93 61.95 23.95 100.00% 0.00%
Forward_N6_S1_13 CAGGAATGACACTATCGGTAGTAAG 79 45.74 52.85 57.75 19.08 100.00% 0.00%
Forward_N6_S1_20 CTCAGGGGGCGACATGCAAATGGGA 462 43.33 52.85 65.48 37.21 100.00% 0.00%
Forward_N6_S1_22 AGGGGGCGACATGCAAATGGGACTA 467 41.29 59.63 67.05 37.59 100.00% 0.00%
Forward_N6_S1_23 GGGGGCGACATGCAAATGGGACTAT 466 42.32 58.66 65.60 37.08 100.00% 0.00%
Forward_N6_S1_24 GGGCGACATGCAAATGGGACTATAC 468 44.67 56.21 62.89 38.74 100.00% 0.00%
Forward_N6_S1_30 CGTAAGTTGGGAAATGGGTCAAGCA 518 40.80 49.13 61.75 30.02 100.00% 0.00%
Forward_N6_S1_39 CCATGACGGCATATCAAGAATGTCA 596 47.90 56.27 59.93 39.57 100.00% 0.00%
Forward_N6_S1_40 CATGACGGCATATCAAGAATGTCAA 597 47.90 55.65 59.06 39.28 100.00% 0.00%
Forward_N6_S1_41 ATGACGGCATATCAAGAATGTCAAT 598 47.28 55.44 58.35 38.10 100.00% 0.00%
Forward_N6_S1_43 GCAAATAGACCAGTAATCACTAT 937 41.30 50.00 54.15 39.61 100.00% 0.00%
Forward_N6_S1_52 GAGCGATTGGGATCATGGTCCTGGC 1401 42.74 55.88 66.20 37.27 100.00% 0.00%
Forward_N6_S1_53 AGCGATTGGGATCATGGTCCTGGCA 1402 42.61 56.80 67.58 30.21 100.00% 0.00%
Forward_N6_S1_54 GCGATTGGGATCATGGTCCTGGCAT 1403 46.25 56.06 66.29 32.93 100.00% 0.00%
Forward_N6_S1_59 GTCCTGGCATGATGGTGCTGAAATC 1418 47.13 52.07 63.28 38.34 100.00% 0.00%
Forward_N6_S1_60 CCTGGCATGATGGTGCTGAAATCAT 1417 45.29 52.63 64.28 35.03 100.00% 0.00%
Forward_N6_S1_61 TGGCATGATGGTGCTGAAATCATCT 1422 49.48 55.52 60.94 37.12 100.00% 0.00%
Forward_N6_S1_62 GGCATGATGGTGCTGAAATCATCTA 1423 47.30 53.55 58.96 37.00 100.00% 0.00%
Forward_N6_S1_63 CATGATGGTGCTGAAATCATCTACT 1425 45.99 52.80 58.60 22.48 100.00% 0.00%
Forward_N6_S1_64 ATGATGGTGCTGAAATCATCTACTT 1426 45.62 52.47 58.27 24.11 100.00% 0.00%
Forward_N6_S1_65 TGATGGTGCTGAAATCATCTACTTT 1426 44.50 50.60 56.69 26.49 100.00% 0.00%
Forward_N6_S1_66 GATGGTGCTGAAATCATCTACTTTA 1423 43.46 50.33 56.63 21.28 100.00% 0.00%
Generate primers time: 0
Running process for reverse primers
Build tables time: 0
Reverse_N6_S1_1 ACTTAAAGTAGATGATTTCAGCA 1412 46.66 51.19 52.61 25.45 100.00% 0.00%
Reverse_N6_S1_2 ACTTAAAGTAGATGATTTCAGCACC 1407 48.79 53.24 56.85 23.80 100.00% 0.00%
Reverse_N6_S1_3 CTTAAAGTAGATGATTTCAGCACCA 1406 48.26 52.83 56.33 21.99 100.00% 0.00%
Reverse_N6_S1_4 TTAAAGTAGATGATTTCAGCACCAT 1405 47.05 51.67 55.46 21.86 100.00% 0.00%
Reverse_N6_S1_5 TAAAGTAGATGATTTCAGCACCATC 1404 47.98 52.52 56.19 27.60 100.00% 0.00%
Reverse_N6_S1_6 AAAGTAGATGATTTCAGCACCATCA 1403 50.38 55.14 58.18 30.74 100.00% 0.00%
Reverse_N6_S1_7 AAGTAGATGATTTCAGCACCATCAT 1402 50.25 55.44 58.39 28.17 100.00% 0.00%
Reverse_N6_S1_8 TAGATGATTTCAGCACCATCATGCC 1393 52.18 57.68 60.15 38.83 100.00% 0.00%
Reverse_N6_S1_13 CATCATGCCAGGACCATGATCCCAA 1383 47.09 56.11 63.78 35.58 100.00% 0.00%
Reverse_N6_S1_14 ATCATGCCAGGACCATGATCCCAAT 1382 49.10 56.04 63.70 32.64 100.00% 0.00%
Reverse_N6_S1_15 TCATGCCAGGACCATGATCCCAATC 1377 45.16 54.29 62.06 32.48 100.00% 0.00%
Reverse_N6_S1_16 CATGCCAGGACCATGATCCCAATCG 1380 49.79 58.53 65.91 29.92 100.00% 0.00%
Reverse_N6_S1_17 TGCCAGGACCATGATCCCAATCGCT 1378 43.42 58.15 66.50 37.73 100.00% 0.00%
Reverse_N6_S1_18 GCCAGGACCATGATCCCAATCGCTC 1377 44.08 58.64 66.52 29.81 100.00% 0.00%
Reverse_N6_S1_26 CTCCTGAGTATCCCGACCAGTTTTG 1221 42.61 51.39 61.72 39.11 100.00% 0.00%
Reverse_N6_S1_31 CCAGGATCTGGGTTTCCTCCTGTTA 1042 40.49 50.11 63.58 27.12 100.00% 0.00%
Reverse_N6_S1_43 CTATAGTGATTACTGGTCTATTTGC 915 42.31 48.49 55.55 38.88 100.00% 0.00%
Reverse_N6_S1_50 ATATTGACATTCTTGATATGCCGTC 576 40.23 48.33 57.06 39.63 100.00% 0.00%
Reverse_N6_S1_54 ATGGGCTGGGTGCTTGACCCATTTC 504 42.48 54.27 66.64 32.90 100.00% 0.00%
Reverse_N6_S1_55 TGGGCTGGGTGCTTGACCCATTTCC 503 46.14 55.54 66.74 28.49 100.00% 0.00%
Reverse_N6_S1_56 GGGCTGGGTGCTTGACCCATTTCCC 503 48.50 57.94 68.79 32.34 100.00% 0.00%
Reverse_N6_S1_57 GGCTGGGTGCTTGACCCATTTCCCA 501 47.45 56.14 67.43 30.68 100.00% 0.00%
Reverse_N6_S1_58 TGGGTGCTTGACCCATTTCCCAACT 498 48.62 56.81 64.91 27.28 100.00% 0.00%
Reverse_N6_S1_59 GGGTGCTTGACCCATTTCCCAACTT 497 49.62 59.71 64.56 25.41 100.00% 0.00%
Reverse_N6_S1_60 GGTGCTTGACCCATTTCCCAACTTA 496 46.08 58.37 62.34 26.51 100.00% 0.00%
Reverse_N6_S1_62 TCCTATCATGTATAGTCCCATTTGC 453 44.46 50.84 56.62 39.48 100.00% 0.00%
Reverse_N6_S1_65 ATGTATAGTCCCATTTGCATGTCGC 446 46.36 53.81 60.98 32.68 100.00% 0.00%
Reverse_N6_S1_66 TGTATAGTCCCATTTGCATGTCGCC 443 41.92 52.96 62.24 28.54 100.00% 0.00%
Reverse_N6_S1_67 GTATAGTCCCATTTGCATGTCGCCC 445 43.05 54.13 63.25 30.73 100.00% 0.00%
Reverse_N6_S1_68 GTCCCATTTGCATGTCGCCCC 444 42.59 53.89 64.94 36.73 100.00% 0.00%
Reverse_N6_S1_73 ACAGGATGTGCCATGAGTTTACTTC 297 40.36 50.63 60.89 33.57 100.00% 0.00%
Reverse_N6_S1_74 AGGATGTGCCATGAGTTTACTTCAC 295 47.37 55.46 60.65 30.77 100.00% 0.00%
Reverse_N6_S1_75 GGATGTGCCATGAGTTTACTTCACA 294 45.25 54.94 60.59 30.80 100.00% 0.00%
Reverse_N6_S1_76 TGTGCCATGAGTTTACTTCACATAG 289 42.28 50.64 58.45 29.96 100.00% 0.00%
Reverse_N6_S1_82 GGCTTACTACCGATAGTGTCATTCC 57 40.25 47.92 60.04 21.73 100.00% 0.00%
Reverse_N6_S1_83 GCTTACTACCGATAGTGTCATTCCT 56 41.85 49.14 59.19 21.22 100.00% 0.00%
Reverse_N6_S1_84 CTTACTACCGATAGTGTCATTCCTG 55 44.41 50.75 58.24 26.84 100.00% 0.00%
Reverse_N6_S1_85 TTACTACCGATAGTGTCATTCCTGT 54 43.49 50.00 57.52 24.46 100.00% 0.00%
Reverse_N6_S1_86 CTACCGATAGTGTCATTCCTGTAGC 51 40.88 49.23 59.55 31.00 100.00% 0.00%
Reverse_N6_S1_87 ACCGATAGTGTCATTCCTGTAGCTG 49 45.41 52.79 61.60 30.19 100.00% 0.00%
Reverse_N6_S1_88 CCGATAGTGTCATTCCTGTAGCTGA 48 41.51 49.22 60.70 29.19 100.00% 0.00%
Reverse_N6_S1_93 GCTGAAATGCATATTATCTTCTGAT 28 41.79 50.95 55.80 38.30 100.00% 0.00%
Reverse_N6_S1_95 ATGCATATTATCTTCTGATTTG 25 41.57 49.88 50.49 39.53 100.00% 0.00%
Generate primers time: 0
Forward_N6_S1_20 CTCAGGGGGCGACATGCAAATGGGA 462 43.33 52.85 65.48 37.21 100.00% 0.00% - Reverse_N6_S1_50 ATATTGACATTCTTGATATGCCGTC 576 40.23 48.33 57.06 39.63 100.00% 0.00% - 100.00% 0.00%
Forward_N6_S1_22 AGGGGGCGACATGCAAATGGGACTA 467 41.29 59.63 67.05 37.59 100.00% 0.00% - Reverse_N6_S1_50 ATATTGACATTCTTGATATGCCGTC 576 40.23 48.33 57.06 39.63 100.00% 0.00% - 100.00% 0.00%
Forward_N6_S1_22 AGGGGGCGACATGCAAATGGGACTA 467 41.29 59.63 67.05 37.59 100.00% 0.00% - Reverse_N6_S1_60 GGTGCTTGACCCATTTCCCAACTTA 496 46.08 58.37 62.34 26.51 100.00% 0.00% - 100.00% 0.00%
Forward_N6_S1_23 GGGGGCGACATGCAAATGGGACTAT 466 42.32 58.66 65.60 37.08 100.00% 0.00% - Reverse_N6_S1_60 GGTGCTTGACCCATTTCCCAACTTA 496 46.08 58.37 62.34 26.51 100.00% 0.00% - 100.00% 0.00%
Forward_N6_S1_24 GGGCGACATGCAAATGGGACTATAC 468 44.67 56.21 62.89 38.74 100.00% 0.00% - Reverse_N6_S1_50 ATATTGACATTCTTGATATGCCGTC 576 40.23 48.33 57.06 39.63 100.00% 0.00% - 100.00% 0.00%
Forward_N6_S1_30 CGTAAGTTGGGAAATGGGTCAAGCA 518 40.80 49.13 61.75 30.02 100.00% 0.00% - Reverse_N6_S1_50 ATATTGACATTCTTGATATGCCGTC 576 40.23 48.33 57.06 39.63 100.00% 0.00% - 100.00% 0.00%
Forward_N6_S1_43 GCAAATAGACCAGTAATCACTAT 937 41.30 50.00 54.15 39.61 100.00% 0.00% - Reverse_N6_S1_31 CCAGGATCTGGGTTTCCTCCTGTTA 1042 40.49 50.11 63.58 27.12 100.00% 0.00% - 100.00% 0.00%
Pairs: 7
Statistics
Total candidates: 164
Pass degeneracy 164 (100.00 %)
Pass GC Content 164 (100.00 %)
Pass GC Clamp 164 (100.00 %)
Pass Max Poly-X 164 (100.00 %)
Pass self complementarity 164 (100.00 %)
Pass self melting 164 (100.00 %)
	Failed self melting low 0 (0.00 %)
	Failed self melting high 0 (0.00 %)
Pass targets 98 (59.76 %)
Pass non targets 72 (43.90 %)
Pass all tests 72 (43.90 %)