                                SEQUENCE LISTING

<110> Litterst, Claudia
      Ugozzoli, Luis A.
      Bio-Rad Laboratories, Inc.

<120> Rapid Detection of Mycoplasma
  Contamination in Cell Culture Samples
  

<130> 002558-0880100PC

<140> WO Not yet assigned    
<141> Not yet assigned  

<150> US 61/256,216       
<151> 2010-10-29  

<160> 27

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 26
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic rpoB gene first degenerate primer

<400> 1
gaagawatgc cwtatttaga agatgg                                      26

<210> 2
<211> 32
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic rpoB gene second degenerate primer

<400> 2
ccrttttgac tyttwccacc magtggttgt tg                               32

<210> 3
<211> 2535
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic hybrid polymerase

<400> 3
atgatcctgg atgctgacta catcactgaa gaaggcaaac cggttatccg tctgttcaaa 60
aaagagaacg gcgaatttaa gattgagcat gatcgcacct ttcgtccata catttacgct 120
ctgctgaaag atgattctaa gattgaggaa gttaaaaaaa tcactgctga gcgccatggc 180
aagattgttc gtatcgttga tgcggaaaag gtagaaaaga aatttctggg cagaccaatc 240
accgtgtgga gactgtattt cgaacatcca caagatgttc cgactattcg cgagaaaatt 300
cgcgaacatt ctgcagttgt tgacatcttc gaatacgata ttccatttgc aaagcgttac 360
ctcatcgaca aaggcctgat accaatggag ggcgatgaag aactcaagct cctggcgttc 420
gctatagcaa ccctctatca cgaaggcgaa gagtttggta aaggcccaat tataatgatc 480
agctatgcag atgaagaaga agcaaaggtg attacttgga aaaaaataga tctcccatac 540
gttgaggttg tatcttccga gcgcgagatg attaagcgct ttctcaaaat tatccgcgag 600
aaggatccgg acattatcat tacttataac ggcgactctt ttgacctccc atatctggcg 660
aaacgcgcag aaaaactcgg tattaaactg actatcggcc gtgatggttc cgagccgaag 720
atgcagcgta tcggcgatat gaccgctgta gaagttaagg gtcgtatcca tttcgacctg 780
tatcatgtaa ttcgtcgtac tattaacctc ccgacttaca ctctcgaggc tgtatatgaa 840
gcaatttttg gtaagccgaa ggagaaggta tacgccgatg agattgcaaa ggcgtgggaa 900
accggtgagg gcctcgagcg tgttgcaaaa tactccatgg aagatgcaaa ggcgacttat 960
gaactcggca aagaattctt cccaatggaa gctcagctct ctcgcctggt tggccaacca 1020
ctgtgggatg tttctcgttc ttccaccggt aacctcgtag agtggtttct cctgcgcaaa 1080
gcgtacgaac gcaacgaact ggctccgaac aagccagatg aacgtgagta tgaacgccgt 1140
ctccgcgagt cttacgctgg tggctttgtt aaagagccag aaaagggcct ctgggaaaac 1200
atcgtgtccc tcgattttcg cgctctgtat ccgtctatta tcattaccca caacgtgtct 1260
ccggatactc tcaaccgcga gggctgcaga aactatgatg ttgctccgga agtaggccac 1320
aagttctgca aggacttccc gggctttatt ccgtctctcc tgaaacgtct gctcgatgaa 1380
cgccaaaaga ttaagactaa aatgaaggcg tcccaggatc cgattgaaaa aataatgctc 1440
gactatcgcc aaagagcgat taaaatcctc gcaaactctt attacggcta ttatggctat 1500
gcaaaagcac gctggtactg taaggagtgt gctgagtccg ttactgcttg gggtcgcgaa 1560
tacatcgagt tcgtgtggaa ggagctcgaa gaaaagtttg gctttaaagt tctctacatt 1620
gacactgatg gtctctatgc gactattccg ggtggtaagt ctgaggaaat taagaaaaag 1680
gctctagaat ttgtggatta cattaacgcg aagctcccgg gtctcctgga gctcgaatat 1740
gaaggctttt ataaacgcgg cttcttcgtt accaagaaga aatatgcgct gattgatgaa 1800
gaaggcaaaa ttattactcg tggtctcgag attgtgcgcc gtgattggag cgaaattgcg 1860
aaagaaactc aagctagagt tctcgaggct attctcaaac acggcaacgt tgaagaagct 1920
gtgagaattg taaaagaagt aacccaaaag ctctctaaat atgaaattcc gccagagaag 1980
ctcgcgattt atgagcagat tactcgcccg ctgcatgagt ataaggcgat tggtccgcac 2040
gtggctgttg caaagagact ggctgctaaa ggcgtgaaaa ttaaaccggg tatggtaatt 2100
ggctacattg tactccgcgg cgatggtccg attagcaacc gtgcaattct agctgaggaa 2160
tacgatccga gaaagcacaa gtatgacgca gaatattaca ttgagaacca ggtgctcccg 2220
gcggtactcc gtattctgga gggttttggc taccgtaagg aagacctccg ctggcaaaag 2280
actaaacaga ctggcctcac ttcttggctc aacattaaaa aatccggtac cggcggtggc 2340
ggtgcaaccg taaagttcaa gtacaaaggc gaagaaaaag aggtagacat ctccaagatc 2400
aagaaagtat ggcgtgtggg caagatgatc tccttcacct acgacgaggg cggtggcaag 2460
accggccgtg gtgcggtaag cgaaaaggac gcgccgaagg agctgctgca gatgctggag 2520
aagcagaaaa agtga                                                  2535

<210> 4
<211> 844
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic hybrid polymerase

<400> 4
Met Ile Leu Asp Ala Asp Tyr Ile Thr Glu Glu Gly Lys Pro Val Ile
 1               5                  10                  15      
Arg Leu Phe Lys Lys Glu Asn Gly Glu Phe Lys Ile Glu His Asp Arg
            20                  25                  30          
Thr Phe Arg Pro Tyr Ile Tyr Ala Leu Leu Lys Asp Asp Ser Lys Ile
        35                  40                  45              
Glu Glu Val Lys Lys Ile Thr Ala Glu Arg His Gly Lys Ile Val Arg
    50                  55                  60                  
Ile Val Asp Ala Glu Lys Val Glu Lys Lys Phe Leu Gly Arg Pro Ile
65                  70                  75                  80  
Thr Val Trp Arg Leu Tyr Phe Glu His Pro Gln Asp Val Pro Thr Ile
                85                  90                  95      
Arg Glu Lys Ile Arg Glu His Ser Ala Val Val Asp Ile Phe Glu Tyr
            100                 105                 110         
Asp Ile Pro Phe Ala Lys Arg Tyr Leu Ile Asp Lys Gly Leu Ile Pro
        115                 120                 125             
Met Glu Gly Asp Glu Glu Leu Lys Leu Leu Ala Phe Ala Ile Ala Thr
    130                 135                 140                 
Leu Tyr His Glu Gly Glu Glu Phe Gly Lys Gly Pro Ile Ile Met Ile
145                 150                 155                 160 
Ser Tyr Ala Asp Glu Glu Glu Ala Lys Val Ile Thr Trp Lys Lys Ile
                165                 170                 175     
Asp Leu Pro Tyr Val Glu Val Val Ser Ser Glu Arg Glu Met Ile Lys
            180                 185                 190         
Arg Phe Leu Lys Ile Ile Arg Glu Lys Asp Pro Asp Ile Ile Ile Thr
        195                 200                 205             
Tyr Asn Gly Asp Ser Phe Asp Leu Pro Tyr Leu Ala Lys Arg Ala Glu
    210                 215                 220                 
Lys Leu Gly Ile Lys Leu Thr Ile Gly Arg Asp Gly Ser Glu Pro Lys
225                 230                 235                 240 
Met Gln Arg Ile Gly Asp Met Thr Ala Val Glu Val Lys Gly Arg Ile
                245                 250                 255     
His Phe Asp Leu Tyr His Val Ile Arg Arg Thr Ile Asn Leu Pro Thr
            260                 265                 270         
Tyr Thr Leu Glu Ala Val Tyr Glu Ala Ile Phe Gly Lys Pro Lys Glu
        275                 280                 285             
Lys Val Tyr Ala Asp Glu Ile Ala Lys Ala Trp Glu Thr Gly Glu Gly
    290                 295                 300                 
Leu Glu Arg Val Ala Lys Tyr Ser Met Glu Asp Ala Lys Ala Thr Tyr
305                 310                 315                 320 
Glu Leu Gly Lys Glu Phe Phe Pro Met Glu Ala Gln Leu Ser Arg Leu
                325                 330                 335     
Val Gly Gln Pro Leu Trp Asp Val Ser Arg Ser Ser Thr Gly Asn Leu
            340                 345                 350         
Val Glu Trp Phe Leu Leu Arg Lys Ala Tyr Glu Arg Asn Glu Leu Ala
        355                 360                 365             
Pro Asn Lys Pro Asp Glu Arg Glu Tyr Glu Arg Arg Leu Arg Glu Ser
    370                 375                 380                 
Tyr Ala Gly Gly Phe Val Lys Glu Pro Glu Lys Gly Leu Trp Glu Asn
385                 390                 395                 400 
Ile Val Ser Leu Asp Phe Arg Ala Leu Tyr Pro Ser Ile Ile Ile Thr
                405                 410                 415     
His Asn Val Ser Pro Asp Thr Leu Asn Arg Glu Gly Cys Arg Asn Tyr
            420                 425                 430         
Asp Val Ala Pro Glu Val Gly His Lys Phe Cys Lys Asp Phe Pro Gly
        435                 440                 445             
Phe Ile Pro Ser Leu Leu Lys Arg Leu Leu Asp Glu Arg Gln Lys Ile
    450                 455                 460                 
Lys Thr Lys Met Lys Ala Ser Gln Asp Pro Ile Glu Lys Ile Met Leu
465                 470                 475                 480 
Asp Tyr Arg Gln Arg Ala Ile Lys Ile Leu Ala Asn Ser Tyr Tyr Gly
                485                 490                 495     
Tyr Tyr Gly Tyr Ala Lys Ala Arg Trp Tyr Cys Lys Glu Cys Ala Glu
            500                 505                 510         
Ser Val Thr Ala Trp Gly Arg Glu Tyr Ile Glu Phe Val Trp Lys Glu
        515                 520                 525             
Leu Glu Glu Lys Phe Gly Phe Lys Val Leu Tyr Ile Asp Thr Asp Gly
    530                 535                 540                 
Leu Tyr Ala Thr Ile Pro Gly Gly Lys Ser Glu Glu Ile Lys Lys Lys
545                 550                 555                 560 
Ala Leu Glu Phe Val Asp Tyr Ile Asn Ala Lys Leu Pro Gly Leu Leu
                565                 570                 575     
Glu Leu Glu Tyr Glu Gly Phe Tyr Lys Arg Gly Phe Phe Val Thr Lys
            580                 585                 590         
Lys Lys Tyr Ala Leu Ile Asp Glu Glu Gly Lys Ile Ile Thr Arg Gly
        595                 600                 605             
Leu Glu Ile Val Arg Arg Asp Trp Ser Glu Ile Ala Lys Glu Thr Gln
    610                 615                 620                 
Ala Arg Val Leu Glu Ala Ile Leu Lys His Gly Asn Val Glu Glu Ala
625                 630                 635                 640 
Val Arg Ile Val Lys Glu Val Thr Gln Lys Leu Ser Lys Tyr Glu Ile
                645                 650                 655     
Pro Pro Glu Lys Leu Ala Ile Tyr Glu Gln Ile Thr Arg Pro Leu His
            660                 665                 670         
Glu Tyr Lys Ala Ile Gly Pro His Val Ala Val Ala Lys Arg Leu Ala
        675                 680                 685             
Ala Lys Gly Val Lys Ile Lys Pro Gly Met Val Ile Gly Tyr Ile Val
    690                 695                 700                 
Leu Arg Gly Asp Gly Pro Ile Ser Asn Arg Ala Ile Leu Ala Glu Glu
705                 710                 715                 720 
Tyr Asp Pro Arg Lys His Lys Tyr Asp Ala Glu Tyr Tyr Ile Glu Asn
                725                 730                 735     
Gln Val Leu Pro Ala Val Leu Arg Ile Leu Glu Gly Phe Gly Tyr Arg
            740                 745                 750         
Lys Glu Asp Leu Arg Trp Gln Lys Thr Lys Gln Thr Gly Leu Thr Ser
        755                 760                 765             
Trp Leu Asn Ile Lys Lys Ser Gly Thr Gly Gly Gly Gly Ala Thr Val
    770                 775                 780                 
Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp Ile Ser Lys Ile
785                 790                 795                 800 
Lys Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe Thr Tyr Asp Glu
                805                 810                 815     
Gly Gly Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys Asp Ala Pro
            820                 825                 830         
Lys Glu Leu Leu Gln Met Leu Glu Lys Gln Lys Lys
        835                 840                 


<210> 5
<211> 23
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic rpoB gene sequencing primer

<400> 5
tgcattttgt catcaaccat gtg                                         23

<210> 6
<211> 17
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic rpoB gene sequencing primer

<400> 6
ccttcacgta tgaacat                                                17

<210> 7
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycobacterium pirum forward primer

<400> 7
gaagatatgc cgcatttaga agatggc                                     27

<210> 8
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycobacterium pirum reverse primer

<400> 8
caacaaccat tgggtggtaa atcacaaaat ggt                              33

<210> 9
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Acholeplasma laidlawii forward primer

<400> 9
gaagatatgc catatttagc agatgga                                     27

<210> 10
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Acholeplasma laidlawii reverse primer

<400> 10
caacaaccaa tgggtggtaa agctcaaaac ggt                              33

<210> 11
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma arginini forward primer

<400> 11
gaagaaatgc catacttaga agatgga                                     27

<210> 12
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma arginini reverse primer

<400> 12
caacaaccac ttggtggtaa atcacaaaat ggt                              33

<210> 13
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma orale forward primer

<400> 13
gaagaaatgc cttatttaga agatggt                                     27

<210> 14
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma orale reverse primer

<400> 14
caacaacctc ttggtggtaa aagtcaaaac ggt                              33

<210> 15
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma salivarium forward primer

<400> 15
gaagatatgc cttatttaga agatgga                                     27

<210> 16
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma salivarium reverse primer

<400> 16
caacaaccac ttggtggaaa gagtcaaaat ggt                              33

<210> 17
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma hominis forward primer

<400> 17
gaagaaatgc cttatttagc agatgga                                     27

<210> 18
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma hominis reverse primer

<400> 18
caacaaccac ttggtggaaa gagtcaaaat ggt                              33

<210> 19
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma fermentans forward primer

<400> 19
gaagatatgc cttatttaga agatggt                                     27

<210> 20
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma fermentans reverse primer

<400> 20
caacaacctc ttggaggtaa gagtcaaaac ggt                              33

<210> 21
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma hyorhinis forward primer

<400> 21
gaagatatgc catttttaga agatgga                                     27

<210> 22
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Mycoplasma hyorhinis reverse primer

<400> 22
caacaaccac ttggaggaaa aagtcaaaac ggt                              33

<210> 23
<211> 26
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Forward primer consensus

<400> 23
gaagawatgc cwtatttaga agatgg                                      26

<210> 24
<211> 32
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic Reverse primer consensus

<400> 24
caacaaccac tkggtggwaa ragtcaaaay gg                               32

<210> 25
<211> 64
<212> PRT
<213> Sulfolobus solfataricus

<220> 
<223> DNA-binding protein 7d (Sso7d)

<400> 25
Met Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp
 1               5                  10                  15      
Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe
            20                  25                  30          
Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg Gly Ala Val Ser Glu
        35                  40                  45              
Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu Glu Lys Gln Lys Lys
    50                  55                  60                  


<210> 26
<211> 66
<212> PRT
<213> Sulfolobus acidocaldarius

<220> 
<223> DNA-binding protein 7d (Sac7d)

<400> 26
Met Val Lys Val Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp
 1               5                  10                  15      
Thr Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys Met Val Ser Phe
            20                  25                  30          
Thr Tyr Asp Asp Asn Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys
        35                  40                  45              
Asp Ala Pro Lys Glu Leu Leu Asp Met Leu Ala Arg Ala Glu Arg Glu
    50                  55                  60                  
Lys Lys
65      


<210> 27
<211> 65
<212> PRT
<213> Sulfolobus acidocaldarius

<220> 
<223> DNA-binding protein 7e (Sac7e)

<400> 27
Met Ala Lys Val Arg Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp
 1               5                  10                  15      
Thr Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys Met Val Ser Phe
            20                  25                  30          
Thr Tyr Asp Asp Asn Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys
        35                  40                  45              
Asp Ala Pro Lys Glu Leu Met Asp Met Leu Ala Arg Ala Glu Lys Lys
    50                  55                  60                  
Lys
65  

