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Technology Seminar (29 Nov 2012)

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Technology Seminar

29 November 2012 (Thur)


Room 7-03, 7/F
The Hong Kong Jockey Club Building for Interdisciplinary Research
5 Sassoon Road, Pokfulam, Hong Kong


By

Dr. Richard Fekete

Sr. Manager

  Part 1: Successful RNA Workflow – Ambion Sample Preparation & Analysis Tools  
  Time: 3:00 – 4:00pm  
  Abstract:

RNA analysis provides an abundance of gene expression information and involves many steps. Often overlooked, stabilizing samples is often the most common time for RNA degradation. Purification of high quality RNA and miRNA with efficient DNA removal is the second key step, and can be done using a variety of methods such as magnetic bead based, filter based or organic extraction. Lysate based preparation products enable a rapid method of getting samples ready for analysis without compromising results. Enriching for targets of interest, either through positive selection or negative selection is also important in obtaining quality data.
Accurate, simple and automation friendly methods for qPCR or the construction small RNA or whole transcriptome RNA libraries for sequencing followed by data analysis are the final steps in the RNA analysis workflow.
Experimental data will be presented to address each of these stages in the RNA Sample to Analysis workflow.

 
  Part 2: Single Cell Expression Analysis  
  Time: 4:15 – 5:15pm  
  Abstract:

When analyzing gene expression profiles from large numbers of cells, the average profile may not be a true representation of the many different profiles that could exist in the cell population (ex., in different states of growth, differentiation or activation). The transcriptional variability of individual cells and any insight into the relationship between specific genes gets lost. One aspect of this emerging field that still needs to be developed is data analysis. How to present data from single cell experiments? What are the proper controls and/or normalization methods to be used? Can you confidently identify sub-populations? The data analysis of high sample numbers with a reduced number of targets is not as straightforward as when using many targets with a few samples (i.e. arrays). Performing cluster analysis and displaying data as histograms can mask the identification of sub-populations. Using FACS we sort thousands of single cells at different time points during stem cell differentiation, stabilize their profiles, preamplify 100 selected genes and analyze these by qPCR. This gives the relative copy number for each transcript in a single cell and allows cells to be classified based on expression profiles. In this way, profiles for numerous cells that have undergone the hESC to NSC pathway are defined, which will in turn enable an in-depth analysis of the growth factor dependence of NSCs, the development of serum free media and the scale-up production of pure NSC from hESCs.

 

ALL ARE WELCOME


Kind Reminder: Please take off your lab coat before coming to the seminar.
Co-organized with Life Technologies Limited Life Technologies Limited Logo
Refreshment to be served
For enquiries, please call 2831-5500 or write to This e-mail address is being protected from spambots. You need JavaScript enabled to view it