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Sample Preparation for Microinjection

art to agglutinate leading to blockages of the injection capillaries. Defrosting should be performed as quickly and gently as possible Before loading the capillary, centrifuge material for 15 minutes at 4C (10,000 x g). Chill supernatant or load directly into capillary.

DNA [5] Purification The highest expression level of microinjected plasmid DNA is achieved when the DNA has been purified by CsCl ethidium bromide gradient centrifugation in accordance with the Maniatis protocol [6].

Dissolve DNA in bidist. water after purification.

Buffers, e.g., PBs, are not recommended as this often leads to blockages of the capillaries.

DNA concentrations of 20 g/ml to 200 g/ml for plasmid DNA injection has been recommended [1].

Storage Store at 20C in small aliquots of 510 l.

Defrosting should be performed as quickly and gently as possible.

Before loading the capillary, centrifuge material for 15 minutes at 4C (10,000 x g). Chill supernatant or load directly into capillary.

Repeated centrifugation does not cause damage but is not necessary for each capillary filling when chilled with ice and used within an hour.

RNA Purification Any standard protocol is suitable for purifying RNA solutions [6].


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