| | |||||||
| Register | Search | Today's Posts | Mark Forums Read |
| Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique. |
| | LinkBack | Thread Tools | Display Modes |
|
#1
| |||
| |||
| Hello everyone, We use these columns to do a homemade Merlin/Wizard-type midiprep. (In fact we just reuse the columns after getting rid of the resin.) Unfortunately, we've lost our last of these columns. Promega has stopped selling the type that we use -- they've seemingly redesigned the columns so as to prevent reuse. (They now have the resin collected in a narrowed part of the column toward the bottom, which you are supposed to physically separate in order to later elute... clever devils! Unfortunately, these new columns also work extremely poorly.) The type that we use is perhaps 50 mL total volume, bottom maybe 4mm diameter, bottom opening maybe 2mm diameter (this is important so they fit into the vacuum manifold). They also have a relatively thick plastic frit at the bottom (important so that resin can't get through), which is the full size of the column diameter. Anyone know of a source for these columns? Thanks, Jenny |
|
#2
| |||
| |||
| In article <fqg455$pn9$[Only registered users see links. ]>, Jenny F. <[Only registered users see links. ]> wrote: Just a suggestion: Buy porous polyethylene sheets that are available from many suppliers in various pore sizes; 30-90 micron is most applicable). I.e., [Only registered users see links. ] (Or just reuse top and bottom frits from Qiagen's maxi prep columns; that's what I do). Then all you need is syringes and a cork borer set. Bore out a frit of appropriate diameter and shove it down 3 ml syringe (blunt end of a 5 ml plastic pipette work best for that). Then, if you insist on having a wider extension for larger volumes, your choice is unlimited - a standard luer connection on all syringes fits very tightly into 3 ml syringe - so any syringe barrel (10, 20, 30, 60 ml) can be used. For unrestricted flow through such improvised column extension, it helps to use a cork borer to remove internal luer connection from it. Such a column (3 ml syringe) can be placed into 15 ml Faclon tube and spun at up tp 2,000 rpm in a typical swinging bucket rotor suitable for such tubes. I use this setup and many of its variations routinely for protein purification and for making spin columns for buffer exchange. The parts (3 ml syringes with frits and larger syringes with a widened exit) can be reused basically indefinitely. Alternatively, if your resin volume is < 0.7 ml, all luer lock syringes snag very tightly into Qiagen's pink (but not blue) columns. DK |
| Tags |
| columns , midiprep , plastic , source , wizard or merlin |
| Thread Tools | |
| Display Modes | |
|
|
| | ||||
| Thread | Thread Starter | Forum | Replies | Last Post |
| the absolutely final complete collection of ideas | blochee | Physics Forum | 2 | 06-15-2007 06:31 AM |
| the absolutely final complete collection of ideas | blochee | Physics Forum | 0 | 06-14-2007 10:30 PM |
| RE: Re: affinity columns (Allison) (DK) | Dan Guire | Protein Forum | 0 | 03-30-2007 10:35 PM |
| Time After Time | EL | Physics Forum | 21 | 07-04-2005 06:58 PM |
| Role of RNA Silencing in Plants | Raghavendra | Botany Forum | 2 | 10-17-2003 10:20 AM |