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I am reading a paper and one sentence is the following: Glycocalyx is unmanifest
now, what does it mean?
I think that manifest means evident, so unmanifest must mean not evident? Thus not present?
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Glycocalyx is unmanifest
Yesterday, 11:00 AM
DNA extraction using qiagen kit
Yesterday, 01:25 AM
Hallo all,
I am doing a DNA extraction with a qiagen kit (http://www.qiagen.co...ringinformation).
Now I wonder what the ATL and AL buffers are.
Also the AW1 and AW2 buffer.
What I found so far is:
ATL : animal tissue lysis. This is pretty straight forward and I understand this.
AL: here is where I get confused => I found (googling) its a cell lysis buffer ? But why would I need another lysis buffer? Isnt ATL used to lyse the tissue and cells? or?
AW1: used to denature the proteins so they can pass through the filter
AW2: 70% ethanol to wash the salts out. It also contains a qiagen concentate.. but no idea what that means..
Any ideas about that AL buffer ?
I am doing a DNA extraction with a qiagen kit (http://www.qiagen.co...ringinformation).
Now I wonder what the ATL and AL buffers are.
Also the AW1 and AW2 buffer.
What I found so far is:
ATL : animal tissue lysis. This is pretty straight forward and I understand this.
AL: here is where I get confused => I found (googling) its a cell lysis buffer ? But why would I need another lysis buffer? Isnt ATL used to lyse the tissue and cells? or?
AW1: used to denature the proteins so they can pass through the filter
AW2: 70% ethanol to wash the salts out. It also contains a qiagen concentate.. but no idea what that means..
Any ideas about that AL buffer ?
competent cells
22 May 2013 - 08:03 AM
Dear all,
I always work with competent cells that I get from a stock of electrocompetent cells we purcharged a while ago, however someone is working rubidium chloride competent cells.
Now I wonder how does this work?
I looked a bit and came up with this, from wikipedia:
RbCl transformation for competent cells is arguably the compound’s most abundant use. Cells treated with a hypotonic solution containing RbCl expand. As a result, the expulsion of membrane proteins allows negatively charged DNA to bind.
I wonder what the role of rubidium chloride is in this? I understand the hypotonic part, but what has that to do with rubidium chloride?
I always work with competent cells that I get from a stock of electrocompetent cells we purcharged a while ago, however someone is working rubidium chloride competent cells.
Now I wonder how does this work?
I looked a bit and came up with this, from wikipedia:
RbCl transformation for competent cells is arguably the compound’s most abundant use. Cells treated with a hypotonic solution containing RbCl expand. As a result, the expulsion of membrane proteins allows negatively charged DNA to bind.
I wonder what the role of rubidium chloride is in this? I understand the hypotonic part, but what has that to do with rubidium chloride?
Role of sodium acetate in DNA extraction/precipitation
22 May 2013 - 06:07 AM
Dear all,
I got a bit confused about the role of sodium acetate in DNA extraction protocols.
I was told it helps to neutralise the DNA charge so that DNA could be precipitated easier with the ethanol added.
However I also saw on some webpages that it also helps precipitng the DNA by binding to proteins (bound to the DNA).
I can understand it helps precipitating the DNA, but if it precipitates with the DNA + proteins , than how do you get rid of those protains after the precipitation?
Because you simple "clean" it with ethanol to remove salts, but you dont remove the proteins , or do you?
I got a bit confused about the role of sodium acetate in DNA extraction protocols.
I was told it helps to neutralise the DNA charge so that DNA could be precipitated easier with the ethanol added.
However I also saw on some webpages that it also helps precipitng the DNA by binding to proteins (bound to the DNA).
I can understand it helps precipitating the DNA, but if it precipitates with the DNA + proteins , than how do you get rid of those protains after the precipitation?
Because you simple "clean" it with ethanol to remove salts, but you dont remove the proteins , or do you?
clogging up qiagen filters
29 March 2013 - 10:29 AM
Dear all,
I am used to work with phenol-chloroform extractions, but recentely I tried a qiagen DNA extraction kit.
I am wondering if it is possible that the filters can get clogged up?
What is weird is that I see a big pellet in the recipient tube (the ones you trow away). So the celldebris does manage to go through the filter, but is it possible that the DNA is also "forced" through this filter with the cell debris? Or perhaps the cell debris is clogging up the filters?
I get very very low amounts of DNA.
thanks in advance
I am used to work with phenol-chloroform extractions, but recentely I tried a qiagen DNA extraction kit.
I am wondering if it is possible that the filters can get clogged up?
What is weird is that I see a big pellet in the recipient tube (the ones you trow away). So the celldebris does manage to go through the filter, but is it possible that the DNA is also "forced" through this filter with the cell debris? Or perhaps the cell debris is clogging up the filters?
I get very very low amounts of DNA.
thanks in advance
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