Newcastle University Wetlab/16 September 2008

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(New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Tuesday 16th September== [[Image:16091.jpg|thumb|right|300px|Gel showing pGFPrrnB-ncl08 and pGFPrrnB-BBa_I746107 restrictions from di...)
 
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[[Image:16091.jpg|thumb|right|300px|Gel showing pGFPrrnB-ncl08 and pGFPrrnB-BBa_I746107 restrictions from different colonies.]]
[[Image:16091.jpg|thumb|right|300px|Gel showing pGFPrrnB-ncl08 and pGFPrrnB-BBa_I746107 restrictions from different colonies.]]
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* Plasmid [[isolated]] from pGFPrrnB-ncl08 colonies 7 + 11 and pGFPrrnB-BB107 colonies 3 + 5 ON cultures.
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* Plasmid [[Isolating Plasmid from Cells (Miniprep)|isolated]] from pGFPrrnB-ncl08 colonies 7 + 11 and pGFPrrnB-BB107 colonies 3 + 5 ON cultures.
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* A Sample of each of the 4 isolated plasmids were [[restricted]] using 7.5μl plasmid and 0.5μl each enzyme in 15μl total volume:
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* A Sample of each of the 4 isolated plasmids were [[Restricting Plasmids (Double Restriction)|restricted]] using 7.5μl plasmid and 0.5μl each enzyme in 15μl total volume:
- pGFPrrnB-ncl08 restricted with EcoRI + NheI
- pGFPrrnB-ncl08 restricted with EcoRI + NheI

Latest revision as of 16:44, 28 October 2008

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Newcastle University

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Home >> Wet Lab >> Newcastle Wet Lab Journal

Wet lab work was carried out from 4 August to 19 September, Mondays to Fridays. Please click on a day to see the lab notebook.

August
MTWTFSS
        [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/1_August_2008&action=edit 1] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/2_August_2008&action=edit 2] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/3_August_2008&action=edit 3]
[http://2008.igem.org/Newcastle_University_Wetlab/4_August_2008 4] [http://2008.igem.org/Newcastle_University_Wetlab/5_August_2008 5] [http://2008.igem.org/Newcastle_University_Wetlab/6_August_2008 6] [http://2008.igem.org/Newcastle_University_Wetlab/7_August_2008 7] [http://2008.igem.org/Newcastle_University_Wetlab/8_August_2008 8] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/9_August_2008&action=edit 9] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/10_August_2008&action=edit 10]
[http://2008.igem.org/Newcastle_University_Wetlab/11_August_2008 11] [http://2008.igem.org/Newcastle_University_Wetlab/12_August_2008 12] [http://2008.igem.org/Newcastle_University_Wetlab/13_August_2008 13] [http://2008.igem.org/Newcastle_University_Wetlab/14_August_2008 14] [http://2008.igem.org/Newcastle_University_Wetlab/15_August_2008 15] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/16_August_2008&action=edit 16] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/17_August_2008&action=edit 17]
[http://2008.igem.org/Newcastle_University_Wetlab/18_August_2008 18] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/19_August_2008&action=edit 19] [http://2008.igem.org/Newcastle_University_Wetlab/20_August_2008 20] [http://2008.igem.org/Newcastle_University_Wetlab/21_August_2008 21] [http://2008.igem.org/Newcastle_University_Wetlab/22_August_2008 22] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/23_August_2008&action=edit 23] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/24_August_2008&action=edit 24]
[http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/25_August_2008&action=edit 25] [http://2008.igem.org/Newcastle_University_Wetlab/26_August_2008 26] [http://2008.igem.org/Newcastle_University_Wetlab/27_August_2008 27] [http://2008.igem.org/Newcastle_University_Wetlab/28_August_2008 28] [http://2008.igem.org/Newcastle_University_Wetlab/29_August_2008 29] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/30_August_2008&action=edit 30] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/31_August_2008&action=edit 31]
September
MTWTFSS
[http://2008.igem.org/Newcastle_University_Wetlab/1_September_2008 1] [http://2008.igem.org/Newcastle_University_Wetlab/2_September_2008 2] [http://2008.igem.org/Newcastle_University_Wetlab/3_September_2008 3] [http://2008.igem.org/Newcastle_University_Wetlab/4_September_2008 4] [http://2008.igem.org/Newcastle_University_Wetlab/5_September_2008 5] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/6_September_2008&action=edit 6] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/7_September_2008&action=edit 7]
[http://2008.igem.org/Newcastle_University_Wetlab/8_September_2008 8] [http://2008.igem.org/Newcastle_University_Wetlab/9_September_2008 9] [http://2008.igem.org/Newcastle_University_Wetlab/10_September_2008 10] [http://2008.igem.org/Newcastle_University_Wetlab/11_September_2008 11] [http://2008.igem.org/Newcastle_University_Wetlab/12_September_2008 12] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/13_September_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/14_September_2008&action=edit 14]
[http://2008.igem.org/Newcastle_University_Wetlab/15_September_2008 15] [http://2008.igem.org/Newcastle_University_Wetlab/16_September_2008 16] [http://2008.igem.org/Newcastle_University_Wetlab/17_September_2008 17] [http://2008.igem.org/Newcastle_University_Wetlab/18_September_2008 18] [http://2008.igem.org/Newcastle_University_Wetlab/19_September_2008 19] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/20_September_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/21_September_2008&action=edit 21]
[http://2008.igem.org/Newcastle_University_Wetlab/22_September_2008 22] [http://2008.igem.org/Newcastle_University_Wetlab/23_September_2008 23] [http://2008.igem.org/Newcastle_University_Wetlab/24_September_2008 24] [http://2008.igem.org/Newcastle_University_Wetlab/25_September_2008 25] [http://2008.igem.org/Newcastle_University_Wetlab/26_September_2008 26] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/27_September_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/28_September_2008&action=edit 28]
[http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/29_September_2008&action=edit 29] [http://2008.igem.org/Newcastle_University_Wetlab/30_September_2008 30]

Tuesday 16th September

Gel showing pGFPrrnB-ncl08 and pGFPrrnB-BBa_I746107 restrictions from different colonies.
  • Plasmid isolated from pGFPrrnB-ncl08 colonies 7 + 11 and pGFPrrnB-BB107 colonies 3 + 5 ON cultures.
  • A Sample of each of the 4 isolated plasmids were restricted using 7.5μl plasmid and 0.5μl each enzyme in 15μl total volume:

- pGFPrrnB-ncl08 restricted with EcoRI + NheI

- pGFPrrnB-BB107 restricted with EcoRI + SpeI

Gel: 

  Expected band sizes:
              * Lane 2: ~ 8.4kb and 2.2kb
              * Lane 3: ~ 8.4kb and 2.2kb
              * Lane 4: ~ 8.4kb and 3.2kb
              * lane 5: ~ 8.4kb and 3.2kb

Lane 1: 1kb ladder

Lane 2: pGFPrrnB-ncl08 colony 7 restricted with EcoR1 and Nhe1

Lane 3: pGFPrrnB-ncl08 colony 11 restricted with EcoR1 and Nhe1

Lane 4: pGFPrrnB-BBa_I746107 colony 3 restricted with EcoR1 and Spe1

lane 5: pGFPrrnB-BBa_I746107 colony 5 restricted with EcoR1 and Spe1

As you can see from our gel, all 4 restrictions yielded the expected sized bands, indicating that the ligations had been successful. The remainder of the unrestricted plasmid from pGFPrrnB-ncl08 colony 7 was stored in the -20°C freezer ready to be sent off for sequencing. The two unrestricted pGFPrrnb-BBa_I746107 plasmid samples were then transformed into B.subtilis 168.

Agar plates made:

- pGFPrrnB-BBa_I746107 colony 3 (chloramphenicol)

- pGFPrrnB-BBa_I746107 colony 5 (chloramphenicol)

- B. subtilis 168 (negative control) (chloamphenicol)

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