IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week10/Chemical and Light

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(New page: =Transformations in Shewie= =Getting Thermoinducible and Lac Inducible GFP into pSC101= =Getting mtrB onto thermo and lac inducible systems=)
(Transformations in Shewie)
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=Transformations in Shewie=
=Transformations in Shewie=
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About 250 ng of DNA added each time.
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==First Set 8/22==
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Plate'''
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| align="center" style="background:#f0f0f0;"|'''Colonies'''
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| align="center" style="background:#f0f0f0;"|'''Size'''
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| align="center" style="background:#f0f0f0;"|'''Color'''
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|-
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| S1 P102 (P108+45) Kan||5 colonies||3mm to 5 mm||Pink
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|-
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| S1 P121+P38 (P124) 1:2.5 Kan A||1 colony||5mm to 7.5mm||Pink
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|-
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| S1 P28 (ColE1) Kan||0 colonies||||
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|-
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| S1 P121+P38(P124) 1:2.5 Kan B||0 colonies||||
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|-
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| S1 P121+P39 1:2.5 Dephos Kan A||0 colonies||||
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|-
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| S1 Topo vector (puc) 2ul Kan X-gal||>50 colonies||1mm||White
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|-
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| S1 Topo vector (puc) 4ul Kan X-gal||>50 colonies||1mm||White
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|-
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| S1 P121+P39 1:2.5 Kan D||0 colonies||||
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|-
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| S1 (-) ctrl (just cells) Kan||0 colonies||||
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|-
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| S1 (+) ctrl (E1 p59b) Kan||2 colonies||5mm ||Pink
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|}
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Poor transformation efficiency in the positive control suggests that there may be a problem with our cells.  All samples, except for P102, were retransformed the following week with fresh cell cultures.
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 +
==Second Set (with new cells) 8/27==
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Plate'''
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| align="center" style="background:#f0f0f0;"|'''Marker'''
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| align="center" style="background:#f0f0f0;"|'''Description'''
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|-
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| S1 P124 1:2.5 A||Kan||14 2-3mm pink colonies, ~100 0.5mm white colonies
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|-
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| S1 P124 1:2.5 B||Kan||2 pink 2mm colonies, >100 0.5mm white colonies
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|-
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| S1 P125 1:2.5 Dephos A||Kan||2 pink 2mm colonies, >100 0.5mm white colonies
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|-
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| S1 P125 1:2.5 D||Kan||8 2mm pink colonies, ~100 0.5mm white colonies
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|-
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| S1 P28||Kan||~8 1-2mm pink colonies, ~50 0.5mm white colonies
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|-
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| S1 TOPO 2uL||Kan||Lawn of (>200) 0.5mm white colonies
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|-
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| S1 TOPO 4 uL (w/ Xgal)||Kan||Lawn of (>200) 0.5mm white colonies
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|-
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| S1 P59b (+) ctrl||Kan||~50 2mm pink colonies, ~50 0.5mm white colonies
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|-
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| S1 (-) ctrl||Kan||50-100 0.5mm white colonies
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|}
=Getting Thermoinducible and Lac Inducible GFP into pSC101=
=Getting Thermoinducible and Lac Inducible GFP into pSC101=
=Getting mtrB onto thermo and lac inducible systems=
=Getting mtrB onto thermo and lac inducible systems=

Revision as of 01:23, 31 August 2008

Contents

Transformations in Shewie

About 250 ng of DNA added each time.

First Set 8/22

Plate Colonies Size Color
S1 P102 (P108+45) Kan5 colonies3mm to 5 mmPink
S1 P121+P38 (P124) 1:2.5 Kan A1 colony5mm to 7.5mmPink
S1 P28 (ColE1) Kan0 colonies
S1 P121+P38(P124) 1:2.5 Kan B0 colonies
S1 P121+P39 1:2.5 Dephos Kan A0 colonies
S1 Topo vector (puc) 2ul Kan X-gal>50 colonies1mmWhite
S1 Topo vector (puc) 4ul Kan X-gal>50 colonies1mmWhite
S1 P121+P39 1:2.5 Kan D0 colonies
S1 (-) ctrl (just cells) Kan0 colonies
S1 (+) ctrl (E1 p59b) Kan2 colonies5mm Pink

Poor transformation efficiency in the positive control suggests that there may be a problem with our cells. All samples, except for P102, were retransformed the following week with fresh cell cultures.

Second Set (with new cells) 8/27

Plate Marker Description
S1 P124 1:2.5 AKan14 2-3mm pink colonies, ~100 0.5mm white colonies
S1 P124 1:2.5 BKan2 pink 2mm colonies, >100 0.5mm white colonies
S1 P125 1:2.5 Dephos AKan2 pink 2mm colonies, >100 0.5mm white colonies
S1 P125 1:2.5 DKan8 2mm pink colonies, ~100 0.5mm white colonies
S1 P28Kan~8 1-2mm pink colonies, ~50 0.5mm white colonies
S1 TOPO 2uLKanLawn of (>200) 0.5mm white colonies
S1 TOPO 4 uL (w/ Xgal)KanLawn of (>200) 0.5mm white colonies
S1 P59b (+) ctrlKan~50 2mm pink colonies, ~50 0.5mm white colonies
S1 (-) ctrlKan50-100 0.5mm white colonies

Getting Thermoinducible and Lac Inducible GFP into pSC101

Getting mtrB onto thermo and lac inducible systems

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