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Typus
Verschleierung
Bearbeiter
Hindemith
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Untersuchte Arbeit:
Seite: 28, Zeilen: 1-9
Quelle: Embark 2004
Seite(n): 48, Zeilen: 17ff
3.3.3 cRNA injection

After storing overnight, oocytes were injected using glass microcapillaries (filled with the required cRNA) mounted in a micromanipulator-controlled microinjector (Figure 7). Precaution was taken so that cRNA was not degraded by RNAases and that the injection capillary was not clogged with small particles. To avoid those problems several procedures were carried out such as using only sterile pipettes, gloves and DEPC treated water for dilution of cRNA. Glass capillaries were pulled using a normal puller. The tip was manually broken under the microscope (diameter of about 10-20 μm), backfilled with paraffin oil to seal the pipette from air and loaded with cRNA by suction (usually 1-2 μl).

2.2.3 cRNA injection

After storing overnight, oocytes were injected using glass microcapillaries (filled with the required cRNA) mounted in a micromanipulator-controlled microinjector (Fig. 13G). Precaution should be taken that cRNA was not contaminated with RNAases and that the injection capillary was not clogged with small particles. To avoid those problems several procedures were carried out such as using only sterile pipettes, gloves and DEPC treated water for dilution of cRNA.

Glass capillaries were pulled using a normal puller. The tip was manually broken under the microscope (diameter of about 10-20 μm), backfilled with paraffin oil to seal the pipette from air and loaded with cRNA by suction (usually 1-2 μl).

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(Hindemith) LieschenMueller

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