9 replies to this topic

[Jamie15]

Hi All,

Just wondering what you all use for Gel extraction.  I used to use Qaigen, but I have found {edit} are much better and cheaper. Anybody else?

[phage434]

The best one is the one you don't use.  I try very hard to design experiments that eliminate gel band isolation.
You can get rid of it almost entirely by using cloning vectors with different resistances, three way ligations, and vector preparation by PCR and DpnI cuts.  My robot is very good at handling liquids, but very bad at cutting out the right band.

[Adrian K]

I used Promega before, now using MEGA-spin™ Agarose Gel Extraction Kit (iNtRON Biotechnology) but it did failed me once due to malfunctioned column...
Agree with phage434, There is no such thing as Best Gel Extraction Kits...

phage434, on 15 June 2011 - 01:53 PM, said:

The best one is the one you don't use.  I try very hard to design experiments that eliminate gel band isolation.
You can get rid of it almost entirely by using cloning vectors with different resistances, three way ligations, and vector preparation by PCR and DpnI cuts.  My robot is very good at handling liquids, but very bad at cutting out the right band.

How much is the price of your robot?

[gyma]

I have been using Wizard® SV Gel and PCR Clean-Up from promega all the time. It works very well. I use it for gel extraction and DNA purification because I dont want to use phenol/chloroform.

[Trof]

I made a miniproject for my student, where we split a reaction in a half and run it on gel, cut out, isolated one with Qiagen MinElute and the other with Geneaid. Eluted  it in the same volume and 260A measurement and gel visualisation both showed Geneaid to have only half of the Qiagen concentration (20 instead of 40ng/ul). We used couple of samples and two different amplicons (smaller around 300 bp, larger 600 bp) and the results were consistent.

It's no serious study, but I will continue to use Qiagen, you could get even more concentrated sample with elution in a smaller volume.
Colleagues told me they don't mind using Geneaid, because they get enough of DNA for sequencing and it's cheaper, but as I was using Geneaid kit for plasmid isolation few days ago, I noticed their columns were poorly manufactured, with excess plastics on the edges and even some plastic "hair" inside one column.

[perneseblue]

Adrian K, on 15 June 2011 - 06:44 PM, said:

phage434, on 15 June 2011 - 01:53 PM, said:

The best one is the one you don't use.  I try very hard to design experiments that eliminate gel band isolation.
You can get rid of it almost entirely by using cloning vectors with different resistances, three way ligations, and vector preparation by PCR and DpnI cuts.  My robot is very good at handling liquids, but very bad at cutting out the right band.

How much is the price of your robot?

Good one there

[phage434]

We got a deal -- as with most of our equipment -- and paid about $15K for a $150K robot.  Not free, but at 10% of retail.  It's amazing what people throw away and sell at huge discounts in bankruptcy.  We love venture capital funding of start ups.  Other people's start ups.

[Adrian K]

WOW phage434, you are brilliant!  Salute!

I remember months back a company was selling their  "demo unit" of thermal cycler (Piko thermalcycler) at 1/3 of the retail price due to their termination of the distributorship. The usage was around 20 times only, tested and working very well, new, nice and sexy.

However, my supervisor refused to buy it even though I beg her very very hard. FYI, we don't even have a PCR machine in our lab. I become a "graduate begger" begging the whole department when I want to do my PCR. I hold my 96-well plate looking machines one after another as a previously booked machines was somehow being used by their own lab members. You guys got no idea the humiliations I suffered and those sarcastic words I heard all these while from those "rich" labs in my department; they never borrow their machines either in the end. My supervisor don't even care  about all these when I told her my difficulties. When there is publication, she want her names there, and put in not related authors.

I wish I got phage434 as my supervisor, really.

[hobglobin]

Try ebay....I once bought a thermal cycler for 100 € there ...an old one without lids, but three blocks and still working for cooling and heating samples....

[perneseblue]

phage434, on 06 September 2011 - 06:30 PM, said:

We got a deal -- as with most of our equipment -- and paid about $15K for a $150K robot.  Not free, but at 10% of retail.  It's amazing what people throw away and sell at huge discounts in bankruptcy.  We love venture capital funding of start ups.  Other people's start ups.

Echoing Adirian K, amazing find . Where did you find this? Do tell, please. ....