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Sample Preparation for Microinjection

As for DNA, RNA is dissolved in bidist. water after purification.

Concentrations of 12 g/ml should be used for mRNA and up to 10 g/ml for total RNA.

Storage Store at 80C in small aliquots of 510 l.

For longer periods, it is advisable to dissolve and store the cleaned RNA in alcohol instead of water.

Defrosting should be performed as quickly and gently as possible.

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

Only centrifuge and use each aliquot once.

Dyes, fluorescent injection markers [1]
Successful fluorescent injection markers used to identify and follow injected cells are: fluorescence labeled dextrans, antibodies, bovine serum albumin.

Dextran marked e.g. with rhodamine of a concentration of 2 g/ml can be detected for up to 48 hours in the cell.

Fluorescein fades with time and results in harmful radicals, therefore markers should preferably be labeled with rhodamine.

Storage Store at 80C in small aliquots of 510 l.

Many solutions are sensitive to light. Thus, exposure to direct light should be avoided.

To avoid blocking of the capillary during microinjection, solutions containing the markers should be filtered with a syringe filter (pore size 0.2 m) whenever possible. Also, before loading the capillary, the sample should be centrifuged for 15 minutes at 4C (10,000 x g).


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