1 storing and preparing samples and reagents, 2 priming and loading the chip – Bio-Rad Experion DNA Analysis Kits User Manual

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2.1 Storing and Preparing Samples and Reagents

Store all Experion

reagents at 4°C when not in use. Do not store reagents at room temperature for

>2 hr, as this will shorten their shelf life.

Before use, allow all kit reagents to equilibrate to room temperature (~15–20 min). Once thawed, gently
vortex all kit reagents before use. Before opening the tubes, quickly centrifuge them to collect solution
to the bottoms of tubes.

If the DNA gel has frozen, discard it.

Protect the DNA stain and gel-stain solution (GS) from light: store these solutions in a dark place and
keep them covered with foil when using them.

The DNA stain contains DMSO, which is hygroscopic. Cap tightly.

Use prepared GS for up to 1 month. Discard after 1 month; do not refilter.

Do not use coated or treated pipet tips or microcentrifuge tubes (for example, siliconized polypropylene)
for preparation of kit reagents or samples. Use of treated tips or tubes may cause separation artifacts.

Use DNase-free microcentrifuge tubes, pipet tips, and TE buffer as needed. Do not use autoclaved
water for sample or reagent preparation.

2.2 Priming and Loading the Chip

To avoid contamination, wear gloves and handle chips by the edges. Never touch the glass portions of
the chip.

Load the chip on a benchtop or in the priming station. Never load a chip in the electrophoresis station.

Avoid sources of dust and other contaminants when preparing samples and loading the chip. Foreign
particles in reagents, samples, or the wells of the chip interfere with separation. Remove chips from their
packaging immediately before use.

It may be easier to load the chip on a white background. Tilt the chip to look for bubbles.

Use narrow-bore pipet tips for loading the chip (for example, VWR #87001-688 or Rainin #L-10F).

To avoid introducing air bubbles, do the following (for more help with chip loading, refer to the Experion
Training Video in the Experion software Help section under Contents and Index > Contents >
Appendices > Technical Videos):

n

Insert the pipet tip all the way to the bottom of the chip well when dispensing liquids

(this reduces the possibility of trapping air)

n

Hold the tip vertically, perpendicular to the chip surface. Holding the tip at an angle

may trap air bubbles at the bottom of the well

n

When expelling liquid, dispense slowly and only to the first stop on the pipet. Using the

second stop introduces air and bubbles into the liquid. Reverse pipetting is acceptable

Dislodge bubbles at the bottom of a well with a clean pipet tip, or remove the solution and load it again.

Use a primed and loaded chip within 5 min of loading. When chips are not used within this time,
reagents may evaporate, leading to poor results or a chip performance error.

Fill all the chip wells when running an analysis. Use blank samples (prepared with water instead of
sample) or replicates if necessary. All 16 electrode pins must be in contact with liquid; otherwise, an
IV (current voltage) check failure error will occur.

Experion Automated Electrophoresis System

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