Team:Mexico-UNAM-CINVESTAV/Notebook/Week Six

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'''Prepared miniprep for I20260 and E0240, we are going to work on the characterize the promoter'''
'''Prepared miniprep for I20260 and E0240, we are going to work on the characterize the promoter'''
-
'''and and the cold expresion casstte by RPU's.'''
+
'''and the cold expresion casstte by RPU's.'''
[[Image:Biobricks002.jpg|500px]]
[[Image:Biobricks002.jpg|500px]]
 +
 +
 +
 +
 +
 +
==''Tuesday''==
 +
 +
'''Yesterday's overnight inoculum we did miniprep and double digestion with ''EcoR''I and ''Pst''I'''
 +
 +
'''To confirm the ligation of PCR 4.'''
 +
 +
 +
{| border="1"
 +
|-
 +
|DNA
 +
|10μl
 +
|-
 +
|Tango
 +
|1.8μl
 +
|-
 +
|EcoRI
 +
|1μl
 +
|-
 +
|PstI
 +
|1μl
 +
|-
 +
|H2O
 +
|1.20μl
 +
|-
 +
|Total
 +
|15μl
 +
|}
 +
 +
 +
[[Image:PCR4.gif]]
 +
 +
 +
==''Wednesday''==
 +
 +
'''We ran a agarosa's gel to extract from the band E0240 and I20260.'''
 +
 +
 +
{| border="1"
 +
|-
 +
|DNA
 +
|50μl
 +
|-
 +
|Tango
 +
|9μl
 +
|-
 +
|EcoRI
 +
|5μl
 +
|-
 +
|PstI
 +
|5μl
 +
|-
 +
|H2O
 +
|6μl
 +
|-
 +
|Total
 +
|75μl
 +
|}
 +
 +
[[Image:Biobricks010.jpg|400px]]
 +
 +
'''We cut the agarosa's band and purified with the kit.'''
 +
 +
'''At the same time we ran the PCR's with ''EcoR''I and ''Pst''I
 +
 +
{| border="1"
 +
|-
 +
|DNA
 +
|20μl
 +
|-
 +
|Tango
 +
|3.6μl
 +
|-
 +
|EcoRI
 +
|2μl
 +
|-
 +
|PstI
 +
|2μl
 +
|-
 +
|H2O
 +
|2.4μl
 +
|-
 +
|Total
 +
|30μl
 +
|}
 +
 +
 +
[[Image:PCR5.gif]]
 +
 +
 +
 +
 +
==''Thursday''==
 +
 +
'''We Quantified with nano espectrophotometer the concentrations of PCR's products and the'''
 +
 +
'''Band's purified'''
 +
 +
 +
{| border="1"
 +
|
 +
|ng/μl
 +
|-
 +
|I20260
 +
|1.30
 +
|-
 +
|E0240
 +
|3.80
 +
|-
 +
|PCR 1 E and P
 +
|277.00
 +
|-
 +
|PCR 2 E and P
 +
|257.00
 +
|-
 +
|PCR 3 E and P
 +
|284.40
 +
|-
 +
|PCR 4 E and P
 +
|284.70
 +
|}

Latest revision as of 07:31, 27 October 2010


Week #6

11th October - 15 October 2010

Monday

We kept inoculated cells on LB liquid medium at 37ºC overnight, each containing 30ml.

DSC02090.JPG


Prepared miniprep for I20260 and E0240, we are going to work on the characterize the promoter

and the cold expresion casstte by RPU's.


Biobricks002.jpg



Tuesday

Yesterday's overnight inoculum we did miniprep and double digestion with EcoRI and PstI

To confirm the ligation of PCR 4.


DNA 10μl
Tango 1.8μl
EcoRI 1μl
PstI 1μl
H2O 1.20μl
Total 15μl


PCR4.gif


Wednesday

We ran a agarosa's gel to extract from the band E0240 and I20260.


DNA 50μl
Tango 9μl
EcoRI 5μl
PstI 5μl
H2O 6μl
Total 75μl

Biobricks010.jpg

We cut the agarosa's band and purified with the kit.

At the same time we ran the PCR's with EcoRI and PstI

DNA 20μl
Tango 3.6μl
EcoRI 2μl
PstI 2μl
H2O 2.4μl
Total 30μl


PCR5.gif



Thursday

We Quantified with nano espectrophotometer the concentrations of PCR's products and the

Band's purified


ng/μl
I20260 1.30
E0240 3.80
PCR 1 E and P 277.00
PCR 2 E and P 257.00
PCR 3 E and P 284.40
PCR 4 E and P 284.70
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