How to make nano-frit

http://www.abrf.org/index.cfm/list.msg/abrf/73705

Date: Mon, 10 Mar 2008 09:08:48 -0400
From: "J. Will Thompson"
To: "'ABRF Discussion List'"
Subject: RE: nano column frits

Hi Chris and Brian,

I have also used this material with a slightly different approach to make high-quality nanocolumn frits. Our technique uses Kasil 2130 (also available free in 1 liter), formamide, and a glass wool fiber filter. Make a clean cut on the end of the capillary and place the fiber filter on a hard surface like a lab benchtop. Place 20uL of Kasil 2130 on the glass filter. Pipet 20ul of formamide directly on top of the Kasil. Using gloves so you can grip the capillary well, depress the end of the capillary into the glass filter and rotate back and forth between thumb and forefinger. Lift and depress in a different spot on the wetted filter; repeat 15-20 times. Each time the capillary will make a small "hole" in the filter and the result should be a 0.5 to 1 mm frit in the end of the capillary (no need to trim the end of the capillary afterwards). Heat at 100C for 30 min to cure. I have used this to frit capillaries from 30 um to 150 um in diameter.
One note, the Kasil/formamide mix cures very quickly so use one spot on a filter paper per capillary you wish to frit. Since you are only using 20ul at a time, the free sample should give you about 50,000 capillary frits.

Good luck!

Will

J. Will Thompson, Ph.D.
Proteomics Core Facility
B02 Levine Science Research Center
Institute for Genome Sciences & Policy
Duke University Medical Center
will.thompson@duke.edu

Consumables from Bruker Daltonics On-line at http://tinyurl.com/79nmaub

I have look around for list of consumables from Bruker Daltonics. Perhaps it was my faults for not looking hard enough, but for my records, I keep it here at http://tinyurl.com/79nmaub.

In particular, below is bacteria test standards for MALDI Biotyper information and preparation protocols.
http://tinyurl.com/82bgem7


The highly sophisticated Bruker Bacterial Test Standard (BTS) contains a typical extract of Escherichia coli DH5alpha. In a MALDI-TOF mass spectrum, this extract shows a characteristic peptide and protein profile. Moreover, the extract is spiked with two additional proteins. Thus, the BTS covers an overall mass range from 3.6 to 17 kDa. Each tube contains material for the preparation of about 40 spots.


To prepare for use,
0. Prepare 50:47.5:2.5 H2O:ACN:TFA solution ("standard solvent").
1. Add 50 μl of standard solvent to the pellet and dissolve by pipetting up and down for at least 20 times at room temperature. Avoid foaming of the solution!
2. Incubate the standard for 5 min at room temperature and repeat pipetting up and down for at least 20 times.
3. Centrifuge for 2 min at room temperature (13000 rpm).
4. Prepare aliquots containing 5 μl of dissolved BTS. For this purpose, use micro tubes connected with screw caps (see above).
5. Use only one micro tube for sample preparation. Store the remaining aliquots at -18°C or below.