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Determining optimal digestion time for partial digestion

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#1 dentategyrus



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Posted 18 October 2011 - 11:30 AM

I am trying to determine the optimal digestion time for a partial digestion of T. cruzi genome with Sau3A. I don't have the gel picture but there was a time gradient, meaning different digestion times. There were two ladders, we weren't told what they were but I think one was a cDNA ladder and the other a genomic ladder for T. cruzi. Can anyone help?

#2 Zagami Francesco

Zagami Francesco


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Posted 29 October 2017 - 01:46 AM

Procedure to obtain homogenate of T. cruzi from epimastigote culture useful for coating wells MTP: taken 1 ml of epimastigote pure culture (forms cultured at 28 ºC in LIT liquid medium), and centrifuged in 5 ml Eppendorff tube at 4000 rpm at 4 ºC for 10 min. Discard the supernatant and 1 ml of phosphate buffer saline (PBS) at pH7.2 is added, and the tube vortexed. This washing operation is repeated for three times. Then, 1 ml of PBS was added and the tube vortexed. Aliquot and store at -20 °C.

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