Stanford
Genome Technology Center
Technology
Development Group
| Instruments
High Capacity Shaker |
High
Capacity Shaker
We have developed a high capacity, atmosphere controlled shaker for growth of e-coli and phage in 96-well microtiter plates. The shaker holds four 14-plate cassettes ( 56 microtiter plates max. capacity) and shakes them in a synchronized manner to eliminate all vibrational forces and moments, which normally would be transferred to the base by the large rotating masses. Each cassette is housed in a sealed tube which is periodically flushed with 100% oxygen gas. This oxygen enrichment has boosted our M13 growth by a factor of five over standard methods. Growth time is approximately 16 hrs and can be set internally in the shaker electronics. All M13 growth at our center is now performed with this shaker. It has been in operation since May 1997. Growth ProtocolsM13 growthOur typical parameters for M13 growth are as follows:
Performance:Typical growth is approx. 2 E12 to 6 E12 pfu/ml based on inoculation with our automated plaque picker. With optimization we have occasionally reached titers of 1 E13 pfu/ml.Cell GrowthTypical parameters for cell growth are identical to M13 growth except:
Performance:Typical growth is approx. 8 E9 to 1 E10 cfu/ml based on starting titers of 10,000 to 100,000 cells. THIS PERFORMANCE IS STRONGLY DEPENDENT ON INITIAL TITER AND OXYGEN LEVELS. If growth is insufficient, try turning oxygen off entirely. You may then slowly add oxygen on subsequent runs to determine the optimum level for your cells and media.The yields from both these growth protocols tend to be three to five times greater than yields from standard growth methods (ie. standard shakers with no oxygen enrichment). This instrument is now available through Genemachines. For information call them at 650-508-1634 or e-mail them. |
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