Problems with biolistics protocol

Kimberley Snowden kcsnowden at
Thu Mar 27 14:59:24 EST 1997

Do you need to resuspend your gold in water?  When I did this I
resuspended in ethanol and then spread the gold on the macrocarrier,
and allowed the ethanol to evaporate off, before assembling everything
and shooting the tissue.  

chend at (Don Chen) wrote:


>We are attempting our hand in transformation of seed embryo callus
>using established protocols.  We are having a problem with the
>resuspension of the gold beads in water.  We find that the beads stick
>to the sides of the wall of the microcentrifuge tubes and refuse to
>make a pellet.  When washing with 100% EtOH, we experience no
>problems.  We only have the problem when we add sterile water.  The
>Bio-Rad people suggested that we should use Tekmar Treff (sp?) tubes,
>we tried with no noticeable improvement.

>People on campus use tungsten beads, and we are in the process of
>trying them to see if we can get a pellet.  We can understand some
>losses, however, it seems hard to imagine how anyone can quantify the
>amount of DNA is being shot into the callus if there are significant
>losses on the sides of the tube.

>What are the experiences of those of you who have tried the biolistics
>approach with gold beads?  Is it possible that the gold is from a bad
>lot?  The tubes we are using are siliconized (US Scientific).  Could
>these have an adverse effect?  One suggestion from the Bio-Rad tech
>service was to use either a swinging bucket centrifuge or one of the
>old style Eppendorf centifuges that mounted the tubes horizontally in
>the rotor.  Could this help us get all of the gold into a pellet?

>Any suggestions would be greatly appreciated.


>Don Chen            *  Standard disclaimers apply here.
>3450 SW Campus Way  *   
>Corvallis, OR 97331 *
>                    *
>541-750-8741        *

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