Forum :: What happened in this plot...?


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This forum is intended for questions about kinetics, Surface Plasmon Resonance and the instruments related to these techniques.

TOPIC: What happened in this plot...?

What happened in this plot...? 20 Jun 2017 04:02 #1

h1cb68f7.jpg

(linker)

2.jpg

(DNA)

Linker is different sequence, and DNA is same sequence at all of channel.

Linker 1 TTCTTTCTTCTTCCGCTTCC
Linker 2 GAAAGCAACAAGGATAGGGG
Linker 3 TCACCAATCGCACACTACTC

DNA GAGTAGTGTGCGATTGGTGA

Step is PBS-Streptavidin-PBS-Linker-PBS-3X SSC(+0.1% DW)-DNA-3X SSC(+0.1% DW).
Streptavidin curve is perfectly good, but linker and DNA curve is not so good. especially, DNA.

DNA and linker 3 is perfectly bind partner, but in ch1, the plot goes upper during washinã…Ž... is it problwm of dilution? or other things?
Koreanraichu @ Korea Univ.
born at Seoul, Republic of Korea

What happened in this plot...? 25 Jun 2017 11:08 #2

Dear K.S.

I know this is a late answer, but it is difficult to give you some advice without knowing the instrument you using.
Please inform us the type of instrument you use and explain how it works. For instance, are there 1, 2, or 3 syringe pumps or is the flow by a peristaltic pump? How is the buffer / analyte switching done: is there an injection loop or is it done by injection of the sample in the running buffer (biacore style)? The flow channels, are they in series or in parallel? How is the temperature stabilization done? More information can help us better.

On the other hand, when I look at these figures, it looks like an improvement over the earlier you have posted. Part of the signal increment looks like drift. This means that washing probably will solve this. Make sure you have washed the chip (and flow and injection system) properly before capturing the linker DNA. Then wash the system again thoroughly before injection of the DNA.

Kind regards
Arnoud

What happened in this plot...? 29 Jun 2017 08:21 #3

Dear Arnoud.
I use peristatic pump when I inject sample and buffer, in all of channel. and each buffer and sample stored in tube(buffer:50ml falcon tube, sample : 1.5ml ep-tube), and when buffer changes, I mones tube to tube which tube of peristic pump. sample flows 40ul/min.
all of channel flous like =(parellel), they cannot shuffle when flow on chip.

One the other hand, how can reduce drift?
Koreanraichu @ Korea Univ.
born at Seoul, Republic of Korea
Last Edit: 29 Jun 2017 08:59 by Koreanraichu.
Moderators: Arnoud