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Role of Serum and Glucocoritcoid [sic] inducible Kinase SGK1 in the regulation of glucose transport

von Dr. Sankarganesh Jeyaraj

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[1.] Sj/Fragment 026 01 - Diskussion
Zuletzt bearbeitet: 2016-11-23 20:37:32 WiseWoman
Embark 2004, Fragment, Gesichtet, KomplettPlagiat, SMWFragment, Schutzlevel sysop, Sj

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KomplettPlagiat
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Hindemith
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Yes
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Seite: 26, Zeilen: 1-6
Quelle: Embark 2004
Seite(n): 45, Zeilen: 19ff
e. 1 μg of linearised DNA was added to 1 μl rNTPS (20 nM), 2.5 μl Cap analogue (to prevent the degradation of the 5’ end of the synthesized RNA), 1 μl RNAase inhibitor (to protect the RNA from degradation by RNAase) and 2.4 μl 10 x transcription buffer(s). After mixing, 1 μl of T7 polymerase was added and the n incubated at 37°C for 1 hr. 1 μl DNase was added and the mixture was subsequently shaken for 15 min at 37°C. After addition of 100 μl DEPC-water and 125 μl phenol chloroform, the mixture was centrifuged at 13,000 rpm for 2 min. e. 1 μg of linearised DNA was added to 1 μl rNTPS (20 nM), 2.5 μl Cap analogue (to prevent the degradation of the 5’ end of the synthesized RNA), 1 μl RNAase inhibitor (to protect the RNA from degradation by RNAase) and 2.4 μl 10 x transcription buffer(s).

f. After mixing, 1 μl of T7 polymerase was added and the n incubated at 37°C for 1 h.

g. 1 μl DNase was added and the mixture was subsequently shaken for 15 min at 37°C.

h. After addition of 100 μl DEPC-water and 125 μl phenolchloroform, the mixture was centrifuged at 13,000 rpm for 2 min.

Anmerkungen

The source is not given.

Sichter
(Hindemith) LieschenMueller

[2.] Sj/Fragment 026 18 - Diskussion
Zuletzt bearbeitet: 2016-11-23 20:39:53 WiseWoman
Embark 2004, Fragment, Gesichtet, KomplettPlagiat, SMWFragment, Schutzlevel sysop, Sj

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Bearbeiter
Graf Isolan
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Yes
Untersuchte Arbeit:
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Quelle: Embark 2004
Seite(n): 48, Zeilen: 1-16
3.3.2 Preparation of Xenopus oocytes

An adult female Xenopus laevis frog was submersed in one liter of 3- aminobenzoic acid ethyl ester (0.1%) for about 15-30 min (Figure 6A). After the frog was fully anesthetized it was placed on ice for surgery.

A small abdominal incision (1 cm) was carried out and a segment of ovary was removed (Figure 6B,C). Subsequently the wound was closed with a reabsorbable suture (Figure 6D). The frog was then kept wet and warm by placing it in a cavity filled by a small amount of warm water to avoid drowning and hypothermia.

The ovarial sacs were manually separated into groups of 10-20 oocytes, put into a 15 ml tube and then enzymatically defolliculated by treatment with an OR- 2 (Oocytes-Ringer) (Table 1) solution containing 1-2 mg/ml collagenase A for 2-2.5 h at room temperature (Figure 6E) with gentle agitation. Defolliculation of the oocytes was stopped by washing several times with ND96 (Table 1).This step also removes all detritus permitting oocyte sorting. Oocytes were then sorted using a self-made apparatus (Figure 6F). Only large oocytes (stage V or VI) were selected and stored overnight in a ND96 storage solution at 16°C.

2.2.2 Preparation of oocytes

An adult female Xenopus laevis frog was submersed in one liter of 3-aminobenzoic acid ethyl ester (0.1%) for about 15-30 min (Fig. 13A). After the frog was fully anesthetized it was placed on ice for surgery. A small abdominal incision (1 cm) was carried out and a segment of ovary was removed (Fig. 13B, C). Subsequently the wound was closed with a reabsorbable suture (Fig. 13D). The frog was then kept wet and warm by placing it in a cavity filled by a small amount of warm water to avoid drowning and hypothermia.

The ovarial sacs were manually separated into groups of 10-20 oocytes, put into a 15 ml tube and then enzymatically defolliculated by treatment with an OR-2 (Oocytes-Ringer) solution containing 1-2 mg/ml collagenase A for 2-2.5 h at room temperature (Fig. 13E) with gentle agitation.

Defolliculation of the oocytes was stopped by washing several times with ND96. This step also removes all detritus permitting oocyte sorting. Oocytes were then sorted using a self-made apparatus (Fig. 13F). Only large oocytes (stage V or VI) were selected and stored overnight in a ND96 storage solution at 16°C.

Anmerkungen

Nothing has been marked as a citation.

Sichter
(Graf Isolan) LieschenMueller


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