Team:Newcastle/18 August 2010

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(New page: {{Team:Newcastle/mainbanner}} =''yneA''= ==Aims== Today we aim to extract ''yneA''in the biobrick comapatible vector from yesterday's overnight culture. We aim to check the concentrat...)
 
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=''yneA''=
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=Miniprep of filamentous cell part=
==Aims==  
==Aims==  
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Today we aim to extract ''yneA''in the biobrick comapatible vector from yesterday's overnight culture. We aim to check the concentration from the miniprep with the nanodrop then digest the plasmid with EcoR1 and Nhe1, ligate into GFPrrnb and transform cells to grow on a plate overnight.  
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The aim of the experiment is to prepare stocks of the plasmid DNA containing our filamentous cell part.
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==Materials and Protocol==  
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==Materials and Protocol==
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Please refer to protocols mentioned below for materials required:
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*[[Team:Newcastle/Qiagen_Minipreps|Plasmid extraction protocol]]
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*[[TeamNewcastleNanoDrop_Spectrophotometer|Nanodrop protocol]]
==Results==  
==Results==  
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===Nanodrop results===
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{|border=1
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|-
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!
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!''yneA'' 1
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!''yneA'' 2
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!''yneA'' 3
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!''yneA'' 4 
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|-
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!Concentration of DNA ng/µl
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!247.3
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!233.6
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!456.6
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!140.0
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|}
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'''Table 1''': Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.
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[[Image:Nanodrop1882010.jpeg|350px]]
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'''Figure 1''': Screenshot of ''yneA'' tube 1
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[[Image:Nanodrop18820102.jpeg|350px]]
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'''Figure 2''': Screenshot of ''yneA'' tube 2
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[[Image:Nanodrop18820103.jpeg|350px]]
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'''Figure 3''': Screenshot of ''yneA'' tube 3
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[[Image:Nanodrop18820104.jpeg|350px]]
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'''Figure 4''': Screenshot of ''yneA'' tube 4
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==Discussion==
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The concentration of the different tubes range from 140.0 µl/ml to 456.6 µl/ml. The standard value for miniprep extraction is ~150 µg/ml.
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==Conclusion==
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High DNA concentration was obtained.
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'''Go back to our main [[Team:Newcastle/notebook| Lab book]] page'''
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=Double digest of filamentous cell part, pSB1C3 and pGFP-rrnB=
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We performed double digests of the filamentous cell part and pSB1C3 using EcoRI and SpeI, ready for ligation.
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We also performed double digests of the filamentous cell part and pGFP-rrnB  using EcoRI and NheI, ready for ligation.
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==Materials and Protocol==
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Please refer to [[Team:Newcastle/Restriction_digests|Restriction digests]].
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===Gel===
 
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 00:05, 28 October 2010

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Contents

Miniprep of filamentous cell part

Aims

The aim of the experiment is to prepare stocks of the plasmid DNA containing our filamentous cell part.

Materials and Protocol

Please refer to protocols mentioned below for materials required:

Results

yneA 1 yneA 2 yneA 3 yneA 4
Concentration of DNA ng/µl 247.3 233.6 456.6 140.0

Table 1: Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.

Nanodrop1882010.jpeg Figure 1: Screenshot of yneA tube 1 Nanodrop18820102.jpeg Figure 2: Screenshot of yneA tube 2 Nanodrop18820103.jpeg Figure 3: Screenshot of yneA tube 3 Nanodrop18820104.jpeg Figure 4: Screenshot of yneA tube 4

Discussion

The concentration of the different tubes range from 140.0 µl/ml to 456.6 µl/ml. The standard value for miniprep extraction is ~150 µg/ml.

Conclusion

High DNA concentration was obtained.

Go back to our main Lab book page

Double digest of filamentous cell part, pSB1C3 and pGFP-rrnB

We performed double digests of the filamentous cell part and pSB1C3 using EcoRI and SpeI, ready for ligation.

We also performed double digests of the filamentous cell part and pGFP-rrnB using EcoRI and NheI, ready for ligation.

Materials and Protocol

Please refer to Restriction digests.


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