Coflow Autoloader: Sample and Buffer requirements

Samples

• Samples will be loaded into eppendorf 96-well plates. These are provided for you at the beamline.

• The wells hold a maximum volume of 200 µl

• max. sample volume for SEC-SAXS is 95 µl

• max sample volume for Batch mode is 100 µl

Buffer quantities

Typically the buffer of the capillary sheath fluid (coflow) matches the sample buffer for both batch and SEC-SAXS mode. Alternatively water can be used as the sheath fluid if buffer is limited.

For SEC-SAXS, you will need enough buffer for the SEC column plus the CoFlow sheath fluid. The volume of sheath fluid is approximately equal to the SEC column volume. You will also need buffer for purging the pump and priming the lines, including the HPLC to CoFlow and the capillary, which is about 40 ml total.

To calculate minimum buffer quantities:

SEC-SAXS, per buffer= number of samples x column volume x 2.5 for CoFlow + 40 ml

Batch mode, per buffer = number of samples x 2 mL + 10 ml to prime the CoFlow lines

You will need enough buffer in the schott bottles to immerse the filters on the bottom of the buffer tubing (about 2 cm long/1cm wide) so that you don’t draw in air.