Team:Stanford/Notebook/Lab Work/Week 5

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==7/26 Monday==
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===Laura's Notebook===
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helped Francisco aliquot competent cells (thaw main tube- 1 mL- on ice, transfer 50 uL to each of 20 pre-chilled 0.5 mL microfuge tubes)
 +
 +
 +
==7/27 Tuesday==
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===Laura's notebook===
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redo failed ligation: vary vector/insert ratios
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{|
 +
| || prior recipe || ligation 1 || ligation 2 || ligation 3
 +
|-
 +
| water || none || 11.0 || 7.0 || none
 +
|-
 +
| vector || 5.0 || 2.0 || 2.0 || 2.0
 +
|-
 +
| insert || 12.0 || 4.0 || 8.0 || 15.0
 +
|-
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| 10X buffer || 2.0 || 2.0 || 2.0 || 2.0
 +
|-
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| ligase || 1.0 || 1.0 || 1.0 || 1.0
 +
|}
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*vector=B1006 linearized (cut with EcoRI, XbaI; inserts=GFP or RFP cut with EcoRI, SpeI (6 ligations set up)
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*Francisco transformed these
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 +
 +
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==7/28 Wednesday==
 +
===Laura's Lab Notebook===
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miniprepped (ProMega kit): GFP and B1006 terminator (RFP didn't grow)
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NanoDrop data:
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{|
 +
|sample || 260/280 || 260/230 || ng/uL
 +
|-
 +
|B1006 || 1.61 || 0.95 || 49.4
 +
|-
 +
| GFP || 1.83 || 1.22 || 96.4
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|}
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==7/29 Thursday==
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==7/30 Friday==
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Revision as of 22:51, 29 July 2010

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Contents

7/26 Monday

Laura's Notebook

helped Francisco aliquot competent cells (thaw main tube- 1 mL- on ice, transfer 50 uL to each of 20 pre-chilled 0.5 mL microfuge tubes)


7/27 Tuesday

Laura's notebook

redo failed ligation: vary vector/insert ratios

prior recipe ligation 1 ligation 2 ligation 3
water none 11.0 7.0 none
vector 5.0 2.0 2.0 2.0
insert 12.0 4.0 8.0 15.0
10X buffer 2.0 2.0 2.0 2.0
ligase 1.0 1.0 1.0 1.0
  • vector=B1006 linearized (cut with EcoRI, XbaI; inserts=GFP or RFP cut with EcoRI, SpeI (6 ligations set up)
  • Francisco transformed these


7/28 Wednesday

Laura's Lab Notebook

miniprepped (ProMega kit): GFP and B1006 terminator (RFP didn't grow) NanoDrop data:

sample 260/280 260/230 ng/uL
B1006 1.61 0.95 49.4
GFP 1.83 1.22 96.4



7/29 Thursday

7/30 Friday

Retrieved from "http://2010.igem.org/Team:Stanford/Notebook/Lab_Work/Week_5"