Mail-In Instructions for HT-SAXS data collection at SIBYLS

Sample Format

1. Sample Volume - A consistent volume of 20µL - 30µL throughout the plate. It is very important that the volume be the same in all wells. Please note the volume on the spreadsheet. 

2. Buffer properties - Do not use greater than 2% glycerol, it has been reported that 2% is the maximum benefit. With glycerol the viscosity of the solution increases and bubbles become more of a problem.

3. Concentration - A concentration above 1mg/ml is required.

A specific plate is required for collection. You can order the plate and sealing mat from Corning.

 

- Full skirt 96-well PCR clear plate from Corning.

 

- AxyMat Sealing Mat from Corning.

 

IMPORTANT: Surface tension is pretty high in these plates but the drops will fall out if the plate is dropped on a hard floor. When shipping your plate over night, we recommend securing plate with a rubber band between 4°C ice packs. After purification, flash freezing should also be possible. Slow freezing is nearly the worst thing you can do to your protein and will happen if you put your liquid samples directly on an ice pack previously stored in the -20°C. The more 4°C in the box the better.

Please write your lab's name and shift date on the side of the plate with a permanent marker before shipping.

 

Please refer to Instructions page on the HT-SAXS Mail-in website for more detailed information about sample prep, spreadsheet requirements, and shipping information. 

What Will Happen When Your Sample Arrives

  • You will be notified that the samples have arrived.

  • The samples will be placed in the 4°C fridge.

  • The plate will be spun at 3700 rpm for 10 min at 4°C prior each data collection.

  • The plate will be at 10°C during the data collection.

  • Data will be collected as soon as possible by SIBYLS staff and you will be notified when data is available.

    • What You Will Get Back 

  • You will get back integrated scattering profiles in ASCI format in a results directory for each sample.

  • A series of exposures, in equal sub-second time slices, are taken of each well.

  • You will get a spreadsheet with an initial assessment of the sample from beamline staff and a folder containing screenshots of images of your scattering profiles.

  • An html file providing an automated guidance for how many files to use in the guinier region, buffer matching, Rg and radiation damage.

    • More Information

    We have a great deal of information on the web at

    http://bl1231.als.lbl.gov

    http://bioisis.net

    http://bl1231.als.lbl.gov/forum

    We welcome questions and constructive comments and will do our best to address both.