Daniel Keppler, Athena W. Lin's Cervical Cancer: Methods and Protocols PDF

By Daniel Keppler, Athena W. Lin

ISBN-10: 149392012X

ISBN-13: 9781493920129

ISBN-10: 1493920138

ISBN-13: 9781493920136

Representing the main proper systems and applied sciences supporting the development of the sector of HPV-mediated carcinogenesis of the cervix and different anatomical areas of squamocolumnar transition, similar to the anorectum, penis, and oropharynx, Cervical melanoma: tools and Protocols compiles an in depth choice of functional chapters. the 1st 1/2 the booklet covers HPV varieties, pathogenesis of cervical melanoma (CxCA), prevention, and novel strength drug goals, whereas the second one part explores pathology, genomics, modeling of CxCA, and experimental healing suggestions. Written within the hugely profitable Methods in Molecular Biology sequence layout, chapters comprise introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, easily reproducible laboratory protocols, and tips about troubleshooting and keeping off recognized pitfalls.

Authoritative and important, Cervical melanoma: equipment and Protocols serves as a beneficial source to either bench scientists and clinicians who step into the area of high-risk HPVs and CxCA for the 1st time or those that desire to research novel techniques or extend their toolbox for the examine of CxCA.

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Pipette 10 μl of the sample extract into the master mix. Add 10 μl of the positive control to the positive control well and add 10 μl PCR Grade water to the negative control well. 5. Seal the Multiwell Plate with LightCycler® 480 Sealing Foil. 6. Place the Multiwell Plate in a swinging-bucket centrifuge and balance it with a suitable counterweight. Centrifuge for 2 min at 1,500 × g. 7. Load the Multiwell Plate into the LightCycler® 480 system. 8. Run PCR with the corresponding program on the LightCycler® 480 system (see Note 3).

A reverse line probe assay, can be used for the identification of 28 different HPV genotypes. This assay shows a good clinical performance compared to other commercial tests for HPV genotyping in cervical cell specimens and formalin-fixed material [6, 7]. Furthermore, the LiPA assay is commonly used in epidemiological studies and vaccination trials [8, 9]. Our group has developed a SYBR Green I-based real-time protocol for the SPF10 primers on the LightCycler® 480 to achieve simultaneous amplification and detection of the HPV target [10].

It was also tested whether the addition of GSSG to raft cultures could artificially advance the maturation of immature capsids to mature capsids. This treatment was able to enhance viral DNA encapsidation and infectivity, but only if added to cultures before virions reached the cornified envelope [49]. No significant effect was observed on viral DNA encapsidation if GSSG was added when the virus was already within the cornified layer. However, treatment at this time did significantly reduce the infectivity of virions, suggesting that the maturation process is a highly regulated process and more complicated than the sole addition of an oxidizing environment.

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Cervical Cancer: Methods and Protocols by Daniel Keppler, Athena W. Lin

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