Team:Groningen/9 September 2010

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(New page: <br> Peter <br> Chaplin detection <br> Samples from the dry freezer were prepared using TFA. <br> The samples were diluted with 1 mL Q-H20 and put on gels: <br> Gel 1: chppt=e deltapt=e bp...)
 
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'''Assembly of biobricks in pSB1C3-backbone for submission to parts registry''' ''by Maarten''
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O/N cultures of transformants were mini prepped with Sigma Aldrich mini prep kit and control restrictions were performed.
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Control restrictions with EcoRI and PstI on constructs:
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{| class="wikitable"
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|-
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| 2 µL construct
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|-
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| 2 µL Fast Digest Buffer
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|-
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| 0,5 µL EcoRI Fast Digest
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|-
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| 0,5 µL PstI Fast Digest
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|-
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| 15 µL MQ
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|}
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{| class="wikitable"
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|-
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! Construct
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! Expected band sizes (bp)
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|-
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| pSB1C3-C
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| 2031, 848
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|-
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| pSB1C3-DC
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| 2031, 872
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|-
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| pSB1C3-E
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| 2031, 317
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|-
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| pSB1C3-H
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| 2031, 302
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|-
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| pSB1C3-Sortase
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| 2031, 689
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|-
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| pSB1C3-RFP
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| 2031, 1069
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|}
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[[Image:Controlerestrictie3.jpeg]]
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{| class="wikitable"
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|-
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| 1-3
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| 4-6
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| 7-9
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|-
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| Chp C
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| Chp DC
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| Chp E
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|}
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[[Image:Controlerestrictie4.jpeg]]
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{| class="wikitable"
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|-
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| 1 & 2
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| 4-6
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| 8
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|-
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| Chp H
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| Sortase
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| RFP
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|}
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The gels were not very conclusive but samples in lane 3, 5 and 7 in the first gel and lane 1 and 6 in the second gel were sent for sequencing on [[Team:Groningen/15 September 2010 | 15 September]].
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<br>
<br>
Peter
Peter

Latest revision as of 22:41, 7 October 2010

Assembly of biobricks in pSB1C3-backbone for submission to parts registry by Maarten

O/N cultures of transformants were mini prepped with Sigma Aldrich mini prep kit and control restrictions were performed.

Control restrictions with EcoRI and PstI on constructs:

2 µL construct
2 µL Fast Digest Buffer
0,5 µL EcoRI Fast Digest
0,5 µL PstI Fast Digest
15 µL MQ
Construct Expected band sizes (bp)
pSB1C3-C 2031, 848
pSB1C3-DC 2031, 872
pSB1C3-E 2031, 317
pSB1C3-H 2031, 302
pSB1C3-Sortase 2031, 689
pSB1C3-RFP 2031, 1069

Controlerestrictie3.jpeg

1-3 4-6 7-9
Chp C Chp DC Chp E

Controlerestrictie4.jpeg

1 & 2 4-6 8
Chp H Sortase RFP

The gels were not very conclusive but samples in lane 3, 5 and 7 in the first gel and lane 1 and 6 in the second gel were sent for sequencing on 15 September.


Peter
Chaplin detection
Samples from the dry freezer were prepared using TFA.
The samples were diluted with 1 mL Q-H20 and put on gels:
Gel 1: chppt=e deltapt=e bpte e1pte enpte chste deltaste bste e1ste
Gel 2: enste deltapt=4 bpt=4 e1pt=4 enpt=4 deltast=4 bst=4 ent=4
Gel 3: est=4 deltadt=e bdt=e e1dt=e endt=e 1 2 3
Gel 4: 4 e1dt=e bdt=e e1nt=e e1pt=e bpt=e e1st=e b1st=e

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