During our lab meeting these weeks, I just reported the PNAS paper which was been mentioned in Li-Hung's e-mail. The research group combined sorting technique and MDA amplification together, and made a easier way to see the link between genes thus. This is really exciting to me.
So here, I introduce "MDA" first and expect a follow-up discussion with everyone.
What is MDA?
It's a amplification method which is different from PCR . {Phi}29 polymerase were used instead of Taq polymerase and amplify proceeds in isothermal condition. Procedure is schematically drawn in figure1.
A key feature of MDA method is that amplification takes place not in cycles, but in a continuous isothermal replication. This makes amplification much more consistent in output and allows a 105 fold of amplification result. DNA that has been produced using MDA method can then be used for any purpose.
p.s. I have not figured out how to insert a picture into my text. Readers can link to www.pixnet.net/photo/thermo/56331535 to see fig. 1
Ting-Wen
So here, I introduce "MDA" first and expect a follow-up discussion with everyone.
What is MDA?
It's a amplification method which is different from PCR . {Phi}29 polymerase were used instead of Taq polymerase and amplify proceeds in isothermal condition. Procedure is schematically drawn in figure1.
 | 1. Random hexamer primers anneal at multiple sites 2. {Phi}29 polymerase initiates replication at multiple sites on denatured linear DNA. Once the strands elongated reaches downstream replicated DNA, displacement occurs and generates new single stranded DNA. Thus, newly synthesized single strand can be bound by additional primers. 3. Subsequent priming and strand displacement produce large quantities of high molecular weight, double stranded DNA. |
A key feature of MDA method is that amplification takes place not in cycles, but in a continuous isothermal replication. This makes amplification much more consistent in output and allows a 105 fold of amplification result. DNA that has been produced using MDA method can then be used for any purpose.
p.s. I have not figured out how to insert a picture into my text. Readers can link to www.pixnet.net/photo/thermo/56331535 to see fig. 1
Ting-Wen
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Thanks Ting-Wen
Thanks Ting-Wen for sharing. (So sad +.+ What I have written is gone. I need to repeat again.)about FACS
I think this paper shows that FACS/MDA/PCR could be well-combined for microbiology. Does our team have the capability of doing this kind of research? Which part is most difficult for us? (PCR should be OK and MDA looks not so hard because of "kid". Protocol "B" seems quite suitable. Maybe?) Can we get the single cell from FACS or any other ways? (Do we have this powerful MoFlo machine? or other flow cytometer can do the same thing?) I am truly interested in this combination. Does any PI want to set up it? Please count me in!Hsiao-Pei