Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/12 August 2015"

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=08/12/2015=
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==Colony PCR for M1 SwqAB 1 & 2 and M1-9(T7)==
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{| class="wikitable"
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| 2X Taq MM
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| 6.25 uL
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|-
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| VF Primer
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| 0.625 uL
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|-
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| VR Primer
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| 0.625 uL
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|-
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| Template
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| 1 uL
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|-
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| ddH2O
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| 4 uL
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|}
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{| class="wikitable"
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| 95C
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| 3 min
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|-
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| 95C
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| 30 sec
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|-
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| 72C
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| 25 sec (M1SeqAB)/ 1min 40 sec (M1-9)
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|-
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|
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| x30
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|-
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| 72C
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| 5 min
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|-
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| 12C
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| Hold
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|}
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[[File:ColonyPCR0812.jpg |800px|thumb|left|Lane 3-6: M1-9(T7) Colony 1-4. Lane 7-10: M1SeqAB1 Colony 1-4. Lane 11-14: M1SeqAB2 Colony 1-4]]
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Grew liquid culture for
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* all 4 colonies of M1-9 (T7); they all showed band close to the expected size ~1200-1300bp but they are slightly different so decided to sequence all of them.
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* colonies 1-3 for M1SeqAB1
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* none for M1SeqAB2 since we probably do not need this seq construct
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== BsaI Digestion of M1AB, BC and CA==
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{| class="wikitable"
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|
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| 1AB (553.88 ng/uL)
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| 1BC (1477.18 ng/uL)
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| 1CA (648 ng/ uL)
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|-
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|Custmart
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| 5 uL
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| 5 uL
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| 5 uL
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|-
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| DNA
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| 7.2 uL
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| 2.7 uL
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| 6.2 uL
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|-
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| ddH2O
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| 33.8 uL
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| 38.3 uL
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| 34.8 uL
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|-
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| BsaI
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| 4 uL
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| 4 uL
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| 4 uL
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|}
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50C for 2 hr
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65C for 20 min
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[[File:BsaIDig0812.jpg|500px]]
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Gel purified samples

Latest revision as of 06:45, 13 August 2015

iGEM UCLA




08/12/2015

Colony PCR for M1 SwqAB 1 & 2 and M1-9(T7)

2X Taq MM 6.25 uL
VF Primer 0.625 uL
VR Primer 0.625 uL
Template 1 uL
ddH2O 4 uL
95C 3 min
95C 30 sec
72C 25 sec (M1SeqAB)/ 1min 40 sec (M1-9)
x30
72C 5 min
12C Hold
Lane 3-6: M1-9(T7) Colony 1-4. Lane 7-10: M1SeqAB1 Colony 1-4. Lane 11-14: M1SeqAB2 Colony 1-4

Grew liquid culture for

  • all 4 colonies of M1-9 (T7); they all showed band close to the expected size ~1200-1300bp but they are slightly different so decided to sequence all of them.
  • colonies 1-3 for M1SeqAB1
  • none for M1SeqAB2 since we probably do not need this seq construct

BsaI Digestion of M1AB, BC and CA

1AB (553.88 ng/uL) 1BC (1477.18 ng/uL) 1CA (648 ng/ uL)
Custmart 5 uL 5 uL 5 uL
DNA 7.2 uL 2.7 uL 6.2 uL
ddH2O 33.8 uL 38.3 uL 34.8 uL
BsaI 4 uL 4 uL 4 uL

50C for 2 hr 65C for 20 min

BsaIDig0812.jpg

Gel purified samples