FlowChip protocol for Higher Dep Density

[JPC]

Hello, we've just tried out the new SOLiD Flowchip loading protocol which includes a centrifugation step of the Flowchip. Basically it was a disaster, we followed the protocol to the letter but we have still had a substantial loss of beads during the spin step.

Has anyone had any success with this protocol?

[SeqAA]

What kind of densities did you get with the protocol?
We are usually around 200-230k/panel with the standard protocol.

[pmiguel]

Since the beads are magnetic, why won't a magnet work to hold the beads against the surface of the flow chip? Naively I would think the biggest problem would be the "field line" striations:



But it seems like there must be away to smooth those out?

--
Phillip

[JPC]

We don't have a bead density to report as they all came out of the flowcell during the centrifuge step. The protocol describes loading 400M beads per lane instead of 240M so I am guessing this should get a density of over 300k per panel?

From talking to our FAS the flowcell has a 'lip' that should prevent the beads coming back out of the slide even when the ports are down (see image)...now I'm thinking we may have loaded this upside down but it would be nice to hear if anyone has had success with this before we try again. I'll run some tests today and post the results.

[JPC]

just to update this thread, we did a few spins yesterday with ports up and ports down and we lost beads with both orientations. With ports facing down (as advised in the protocol) the loss was for less but it was still significant so we won't be working with this protocol right now.

JC