Overexpression of multivalent transcription factor, ctcf in hela cell lines

Transformation of pCI-CTCF plasmid was done after receiving this recombinant construct from Dr. Elena M. Klenova, Gene Regulation Laboratory, University of Essex, UK. The plasmid was propagated in E.coli DHS-a competence cell. Following transformation, a transformant containing the recombinant pl...

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Main Author:	Nurul Wahida, Ab. Ghani
Format:	Monograph
Language:	English
Published:	Universiti Sains Malaysia 2008
Subjects:	R Medicine (General)
Online Access:	http://eprints.usm.my/48895/ http://eprints.usm.my/48895/1/NURUL%20W%20AHIDA%20BINTI%20AB.%20GHANI%20-%2024%20pages.pdf

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author	Nurul Wahida, Ab. Ghani
author_facet	Nurul Wahida, Ab. Ghani
author_sort	Nurul Wahida, Ab. Ghani
building	USM Institutional Repository
collection	Online Access
description	Transformation of pCI-CTCF plasmid was done after receiving this recombinant construct from Dr. Elena M. Klenova, Gene Regulation Laboratory, University of Essex, UK. The plasmid was propagated in E.coli DHS-a competence cell. Following transformation, a transformant containing the recombinant plasmid was selected and cultured overnight in LB Broth. The plasmid was then extracted using a plasmid extraction kit QIAprep Spin Miniprep Kit (QIAGEN, Germany). Purified plasmid was further analysed and characterized. The plasmid was then transfected into HeLa cells using Calcium Phosphate method. Following transfection, the HeLa cells were lysed and the supernatant of the lysed cells was analysed using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) followed by Western Blotting using anti CTCF monoclonal antibody. There is a positive expression of exogenous CTCF detected in the lysate compare to the normal HeLa cell lines control. However due to short period of time for this project, transfection-efficiency was not properly estimated. This work established conventional, cheap and reasonable transfection method for future functional assay works in the lab.
first_indexed	2025-11-15T18:16:36Z
format	Monograph
id	usm-48895
institution	Universiti Sains Malaysia
institution_category	Local University
language	English
last_indexed	2025-11-15T18:16:36Z
publishDate	2008
publisher	Universiti Sains Malaysia
recordtype	eprints
repository_type	Digital Repository
spelling	usm-488952021-04-14T07:46:34Z http://eprints.usm.my/48895/ Overexpression of multivalent transcription factor, ctcf in hela cell lines Nurul Wahida, Ab. Ghani R Medicine (General) Transformation of pCI-CTCF plasmid was done after receiving this recombinant construct from Dr. Elena M. Klenova, Gene Regulation Laboratory, University of Essex, UK. The plasmid was propagated in E.coli DHS-a competence cell. Following transformation, a transformant containing the recombinant plasmid was selected and cultured overnight in LB Broth. The plasmid was then extracted using a plasmid extraction kit QIAprep Spin Miniprep Kit (QIAGEN, Germany). Purified plasmid was further analysed and characterized. The plasmid was then transfected into HeLa cells using Calcium Phosphate method. Following transfection, the HeLa cells were lysed and the supernatant of the lysed cells was analysed using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) followed by Western Blotting using anti CTCF monoclonal antibody. There is a positive expression of exogenous CTCF detected in the lysate compare to the normal HeLa cell lines control. However due to short period of time for this project, transfection-efficiency was not properly estimated. This work established conventional, cheap and reasonable transfection method for future functional assay works in the lab. Universiti Sains Malaysia 2008-10 Monograph NonPeerReviewed application/pdf en http://eprints.usm.my/48895/1/NURUL%20W%20AHIDA%20BINTI%20AB.%20GHANI%20-%2024%20pages.pdf Nurul Wahida, Ab. Ghani (2008) Overexpression of multivalent transcription factor, ctcf in hela cell lines. Other. Universiti Sains Malaysia. (Submitted)
spellingShingle	R Medicine (General) Nurul Wahida, Ab. Ghani Overexpression of multivalent transcription factor, ctcf in hela cell lines
title	Overexpression of multivalent transcription factor, ctcf in hela cell lines
title_full	Overexpression of multivalent transcription factor, ctcf in hela cell lines
title_fullStr	Overexpression of multivalent transcription factor, ctcf in hela cell lines
title_full_unstemmed	Overexpression of multivalent transcription factor, ctcf in hela cell lines
title_short	Overexpression of multivalent transcription factor, ctcf in hela cell lines
title_sort	overexpression of multivalent transcription factor, ctcf in hela cell lines
topic	R Medicine (General)
url	http://eprints.usm.my/48895/ http://eprints.usm.my/48895/1/NURUL%20W%20AHIDA%20BINTI%20AB.%20GHANI%20-%2024%20pages.pdf

Overexpression of multivalent transcription factor, ctcf in hela cell lines

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