pdna Search Results


atcc 45151d  (ATCC)
94
ATCC atcc 45151d
Atcc 45151d, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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atcc 209396  (ATCC)
86
ATCC atcc 209396
Atcc 209396, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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45152d 31 nibsc  (ATCC)
94
ATCC 45152d 31 nibsc
45152d 31 Nibsc, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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absidia sp  (ATCC)
90
ATCC absidia sp
MICs of CMT-3 and AMB
Absidia Sp, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hek293 cells  (OriGene)
95
OriGene hek293 cells
MICs of CMT-3 and AMB
Hek293 Cells, supplied by OriGene, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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streptococcus  (ATCC)
93
ATCC streptococcus
MICs of CMT-3 and AMB
Streptococcus, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plasmid pam6  (ATCC)
93
ATCC plasmid pam6
MICs of CMT-3 and AMB
Plasmid Pam6, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plasmid dna  (ATCC)
96
ATCC plasmid dna
Fig. 2. ( a ) HPV detection in clinical samples by a low <t>temperature</t> <t>PCR</t> polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV <t>DNA</t> in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).
Plasmid Dna, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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atcc pta 4405  (ATCC)
93
ATCC atcc pta 4405
Fig. 2. ( a ) HPV detection in clinical samples by a low <t>temperature</t> <t>PCR</t> polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV <t>DNA</t> in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).
Atcc Pta 4405, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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40341 u s pat  (ATCC)
90
ATCC 40341 u s pat
Fig. 2. ( a ) HPV detection in clinical samples by a low <t>temperature</t> <t>PCR</t> polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV <t>DNA</t> in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).
40341 U S Pat, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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deposit as pgrn33  (ATCC)
90
ATCC deposit as pgrn33
Fig. 2. ( a ) HPV detection in clinical samples by a low <t>temperature</t> <t>PCR</t> polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV <t>DNA</t> in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).
Deposit As Pgrn33, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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deposit no atcc 209377  (ATCC)
86
ATCC deposit no atcc 209377
Fig. 2. ( a ) HPV detection in clinical samples by a low <t>temperature</t> <t>PCR</t> polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV <t>DNA</t> in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).
Deposit No Atcc 209377, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


MICs of CMT-3 and AMB

Journal:

Article Title: A Chemically Modified Tetracycline (CMT-3) Is a New Antifungal Agent

doi: 10.1128/AAC.46.5.1447-1454.2002

Figure Lengend Snippet: MICs of CMT-3 and AMB

Article Snippet: The MIC of CMT-3 was 2.0 μg/ml, and the 50% inhibitory concentration was about 1.0 μg/ml. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Organism Source a MIC (μg/ml) of: No. of strains CMT-3 AMB C. albicans ATCC 24433, ATCC 76615, ATCC 18804, ATCC 90028, CI 0.25->8 0.12-4.0 17 C. albidus ATCC 34140 2.0 1.0 1 C. glabrata CI 4.0-8.0 0.5-2.0 2 C. krusei ATCC 6258, CI 4.0->8.0 1.0-2.0 3 C. parapsilosis ATCC 22019, CI >8.0 1.0-2.0 2 C. tropicalis ATCC 750, CI 4.0->8.0 0.5-2.0 3 Absidia sp. CI 4.0 4.0 1 A. flavus CI 2.0 1.0 1 A. fumigatus ATCC 1022 2.0 2.0 1 Cunninghamella sp. CI 8.0 >8.0 1 E. floccosum CI 0.5 >8.0 1 Fonsecaea sp. CI 4.0 >8.0 1 M. canis CI 2.0 0.5 1 M. gypseum CI 1.0 4.0 1 P. boydii CI 0.25 >8.0 1 Penicillium sp. CI 0.5 0.25 1 P. variotii ATCC 22319 1.0 0.5 1 P. verrucosa CI 8.0 >8.0 1 Rhizopus sp. CI 1.0 0.5 1 S. apiospermum CI 0.5 >8.0 1 T. mentagrophytes CI 1.0 4.0 1 T. tonsurans CI 2.0 0.25 1 Tricothecium sp. CI 0.5 >8.0 1 T. rubrum ATCC 10218 0.5 1.0 1 Ulocladium sp. CI 1.0 2.0 1 Open in a separate window a CI, clinical isolate.

Techniques:

Inhibition of fungal viability by CMT-3 and AMB

Journal:

Article Title: A Chemically Modified Tetracycline (CMT-3) Is a New Antifungal Agent

doi: 10.1128/AAC.46.5.1447-1454.2002

Figure Lengend Snippet: Inhibition of fungal viability by CMT-3 and AMB

Article Snippet: The MIC of CMT-3 was 2.0 μg/ml, and the 50% inhibitory concentration was about 1.0 μg/ml. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Organism Source a MIC (μg/ml) of: No. of strains CMT-3 AMB C. albicans ATCC 24433, ATCC 76615, ATCC 18804, ATCC 90028, CI 0.25->8 0.12-4.0 17 C. albidus ATCC 34140 2.0 1.0 1 C. glabrata CI 4.0-8.0 0.5-2.0 2 C. krusei ATCC 6258, CI 4.0->8.0 1.0-2.0 3 C. parapsilosis ATCC 22019, CI >8.0 1.0-2.0 2 C. tropicalis ATCC 750, CI 4.0->8.0 0.5-2.0 3 Absidia sp. CI 4.0 4.0 1 A. flavus CI 2.0 1.0 1 A. fumigatus ATCC 1022 2.0 2.0 1 Cunninghamella sp. CI 8.0 >8.0 1 E. floccosum CI 0.5 >8.0 1 Fonsecaea sp. CI 4.0 >8.0 1 M. canis CI 2.0 0.5 1 M. gypseum CI 1.0 4.0 1 P. boydii CI 0.25 >8.0 1 Penicillium sp. CI 0.5 0.25 1 P. variotii ATCC 22319 1.0 0.5 1 P. verrucosa CI 8.0 >8.0 1 Rhizopus sp. CI 1.0 0.5 1 S. apiospermum CI 0.5 >8.0 1 T. mentagrophytes CI 1.0 4.0 1 T. tonsurans CI 2.0 0.25 1 Tricothecium sp. CI 0.5 >8.0 1 T. rubrum ATCC 10218 0.5 1.0 1 Ulocladium sp. CI 1.0 2.0 1 Open in a separate window a CI, clinical isolate.

Techniques: Inhibition, Control

Fig. 2. ( a ) HPV detection in clinical samples by a low temperature PCR polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV DNA in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).

Journal: Methods in Molecular Biology

Article Title: Diagnosis of Sexually Transmitted Diseases

doi: 10.1007/978-1-61779-937-2

Figure Lengend Snippet: Fig. 2. ( a ) HPV detection in clinical samples by a low temperature PCR polymerase. Footnote : Agarose gel electrophoresis plate showing detection of HPV DNA in 1 m L of crude proteinase K digestate of liquid-based cervicovaginal cytology speci- men from seven patients by a low temperature nested PCR. Each PCR mixture contained forward primer 1 m L, reverse primer 1 m L, PCR master mix 20 m L, template, and water added to fi nal volume 25 m L. Primary PCR primers were MY09/ MY11, and the nested PCR primers GP6/MY11. Primary PCR products were transferred by micro-glass rods. All nested PCR amplicons were validated by DNA sequencing: Lane 1, HPV-16; Lane 2, HPV-16; Lane 3, HPV-39; Lane 4, HPV-68; Lane 5, HPV-54; Lane 6, HPV-59; Lane 7, HPV-54; N, negative control; P, plasmid HPV-16 DNA control; M, molecular ruler. Note : Primary PCR did not show a visible PCR amplicon in 3 of 7 HPV-positive specimens. Thermocycling steps: initial heating at 85 °C for 10 min, followed by 30 cycles, each set at 85 °C for 30 s, 40 °C for 30 s, and 65 °C for 1 min. The fi nal extension was 65 °C for 10 min. (b) HPV detection in clinical samples by a Taq PCR polymerase. Footnote : Comparative parallel pri- mary and nested PCR detection of HPV DNA in 1 m L crude proteinase K digestate from the seven patients as for Fig. 2a , using a Taq DNA polymerase. The Taq PCR protocol: GE r Taq DNA polymerase, 2.5 units in 0.25 m L, was used in each PCR with 10×PCR buffer 2.5 m L, 25 mM MgCl 2 2 m L, 5 mM dNTPs 1 m L, forward primer 1 m L, reverse primer 1 m L, template, and water added to fi nal volume 25 m L. The nested PCR mixture contained the same ingredients. Primary PCR products were transferred to nested PCR tubes by micro-glass rods as described in this chapter. Primary PCR primers were MY09/ MY11 and the nested PCR primers GP6/MY11, as for Fig. 2a . Thermocycling steps were set as preheating for 1 min at 94 °C, followed by 30 cycles, each set at 94 °C for 30 s, 54 °C for 2 min and 72 °C for 1 min. The fi nal extension was 72 °C for 10 min for both primary and nested PCR. Note : Although the Taq PCR failed to detect HPV DNA in four of the seven unpuri fi ed clinical samples known to be positive for HPV, it ampli fi ed pure plasmid HPV-16 DNA quite ef fi ciently (lane P).

Article Snippet: Nested PCR with this DNA polymerase can detect 1–10 copies of plasmid DNA of HPV types-16, -18 or -6B purchased from ATCC ( 15 ) .

Techniques: Agarose Gel Electrophoresis, Nested PCR, DNA Sequencing, Negative Control, Plasmid Preparation, Control, Amplification

Fig. 3. Computer-generated electropherogram showing the sequence of a target HPV DNA. Footnote : A segment of DNA sequencing electropherogram with its ABI computer-generated base-calling results, consisting of the last fi ve bases (3 ¢ CCATT---5 ¢ ) of the MY11 primer-binding site on the right and 56 bases downstream thereof. The triad “AAA” sequence, which is invariably positioned 23 bases downstream of the MY11 primer-binding site, is seen for all known HPV genotypes of clinical relevance.

Journal: Methods in Molecular Biology

Article Title: Diagnosis of Sexually Transmitted Diseases

doi: 10.1007/978-1-61779-937-2

Figure Lengend Snippet: Fig. 3. Computer-generated electropherogram showing the sequence of a target HPV DNA. Footnote : A segment of DNA sequencing electropherogram with its ABI computer-generated base-calling results, consisting of the last fi ve bases (3 ¢ CCATT---5 ¢ ) of the MY11 primer-binding site on the right and 56 bases downstream thereof. The triad “AAA” sequence, which is invariably positioned 23 bases downstream of the MY11 primer-binding site, is seen for all known HPV genotypes of clinical relevance.

Article Snippet: Nested PCR with this DNA polymerase can detect 1–10 copies of plasmid DNA of HPV types-16, -18 or -6B purchased from ATCC ( 15 ) .

Techniques: Generated, Sequencing, DNA Sequencing, Binding Assay

Fig. 5. Sequencing electropherogram of more than one HPV target DNA in specimen. Footnote : When the infection is caused by two HPV genotypes with different amplicon sizes (HPV-16 and HPV-18), one end sequence 3 ¢ CCATT---5 ¢ of the MY11 primer site is positioned ahead of the other, as shown in this mixed HPV DNA sequence tracing. The fi rst T of each end segment is indicated by an arrow. This is due to the fact that the size of the GP6/MY11 nested PCR amplicon of HPV-18 is 3-bp longer than that of HPV-16 ( see Fig. 1 ).

Journal: Methods in Molecular Biology

Article Title: Diagnosis of Sexually Transmitted Diseases

doi: 10.1007/978-1-61779-937-2

Figure Lengend Snippet: Fig. 5. Sequencing electropherogram of more than one HPV target DNA in specimen. Footnote : When the infection is caused by two HPV genotypes with different amplicon sizes (HPV-16 and HPV-18), one end sequence 3 ¢ CCATT---5 ¢ of the MY11 primer site is positioned ahead of the other, as shown in this mixed HPV DNA sequence tracing. The fi rst T of each end segment is indicated by an arrow. This is due to the fact that the size of the GP6/MY11 nested PCR amplicon of HPV-18 is 3-bp longer than that of HPV-16 ( see Fig. 1 ).

Article Snippet: Nested PCR with this DNA polymerase can detect 1–10 copies of plasmid DNA of HPV types-16, -18 or -6B purchased from ATCC ( 15 ) .

Techniques: Sequencing, Infection, Amplification, Nested PCR

Fig. 6. Mixed HPV infection turning into single persistent HPV infection on follow-up. Footnote : Mixed HPV infections are usually transient. If not totally cleared on follow-up, they often turn into a single HPV infection, as the case in this 49-year- old woman with a history of reactive Papanicolaou cytology result. In December, 2009, her cervicovaginal cells were posi- tive for a mixed HPV infection with two superimposing staggering 3 ¢ CCATT---5 ¢ MY11 endings on the right of the upper sequence tracing (the fi rst T of each ending is indicated by an arrow ). A repeat HPV test in June 2010 detected a single HPV-59 infection as demonstrated in the lower sequence tracing. Comparison of the two tracings reveals that a DNA sequence characteristic of that of an HPV-59 was overshadowed in the background of the mixed HPV sequences in the electropherogram recorded 6 months earlier.

Journal: Methods in Molecular Biology

Article Title: Diagnosis of Sexually Transmitted Diseases

doi: 10.1007/978-1-61779-937-2

Figure Lengend Snippet: Fig. 6. Mixed HPV infection turning into single persistent HPV infection on follow-up. Footnote : Mixed HPV infections are usually transient. If not totally cleared on follow-up, they often turn into a single HPV infection, as the case in this 49-year- old woman with a history of reactive Papanicolaou cytology result. In December, 2009, her cervicovaginal cells were posi- tive for a mixed HPV infection with two superimposing staggering 3 ¢ CCATT---5 ¢ MY11 endings on the right of the upper sequence tracing (the fi rst T of each ending is indicated by an arrow ). A repeat HPV test in June 2010 detected a single HPV-59 infection as demonstrated in the lower sequence tracing. Comparison of the two tracings reveals that a DNA sequence characteristic of that of an HPV-59 was overshadowed in the background of the mixed HPV sequences in the electropherogram recorded 6 months earlier.

Article Snippet: Nested PCR with this DNA polymerase can detect 1–10 copies of plasmid DNA of HPV types-16, -18 or -6B purchased from ATCC ( 15 ) .

Techniques: Infection, Sequencing, Comparison