Newcastle University Wetlab/17 September 2008

From 2008.igem.org

(Difference between revisions)
(New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Wednesday 17th September== * Results of ON agar plate cultures: - pGFPrrnB-BBa_I746107 colony 3 (chloramphenicol) = ~100 colonies ...)
 
Line 11: Line 11:
- ''B. subtilis'' 168 (negative control) (chloamphenicol) = O colonies
- ''B. subtilis'' 168 (negative control) (chloamphenicol) = O colonies
-
These colonies show that the BBa_I746107 insert has successfully transformed into the ''B. subtilis'' chromosomal DNA. To avoid confusion between ''E. coli'' and ''B. subtilis'', it was decided to rename the insert from pGFPrrnB-ncl08 (in ''E. coli'') to iGEMBB107 (in ''B. subtilis''). It was important to then check that only this insert had integrated (and not the rest of the vector plasmid) and that it had integrated in the correct place. Therefore the following [[agar]] plate cultures were made:
+
These colonies show that the BBa_I746107 insert has successfully transformed into the ''B. subtilis'' chromosomal DNA. To avoid confusion between ''E. coli'' and ''B. subtilis'', it was decided to rename the insert from pGFPrrnB-ncl08 (in ''E. coli'') to iGEMBB107 (in ''B. subtilis''). It was important to then check that only this insert had integrated (and not the rest of the vector plasmid) and that it had integrated in the correct place. Therefore the following [[Making Agar Plates|agar]] plate cultures were made:
- iGEMBB107 colony 3 (starch + chloramphenicol)
- iGEMBB107 colony 3 (starch + chloramphenicol)

Latest revision as of 16:44, 28 October 2008

Bugbuster-logo-red.png
Ncl uni logo.jpg


Newcastle University

GOLD MEDAL WINNER 2008

Home Team Original Aims Software Modelling Proof of Concept Brick Wet Lab Conclusions


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]

Wednesday 17th September

  • Results of ON agar plate cultures:

- pGFPrrnB-BBa_I746107 colony 3 (chloramphenicol) = ~100 colonies

- pGFPrrnB-BBa_I746107 colony 5 (chloramphenicol) = ~80 colonies

- B. subtilis 168 (negative control) (chloamphenicol) = O colonies

These colonies show that the BBa_I746107 insert has successfully transformed into the B. subtilis chromosomal DNA. To avoid confusion between E. coli and B. subtilis, it was decided to rename the insert from pGFPrrnB-ncl08 (in E. coli) to iGEMBB107 (in B. subtilis). It was important to then check that only this insert had integrated (and not the rest of the vector plasmid) and that it had integrated in the correct place. Therefore the following agar plate cultures were made:

- iGEMBB107 colony 3 (starch + chloramphenicol)

- iGEMBB107 colony 3 (spectinomycin)

- iGEMBB107 colony 3 (chloramphenicol)

- iGEMBB107 colony 5 (starch + chloramphenicol)

- iGEMBB107 colony 5 (spectinomycin)

- iGEMBB107 colony 5 (chloramphenicol)

  • ON cultures were made from glycerol stocks to check for correct insert (in 3mL LB):

- pGFPrrnB-ncl08 colony 7

- pGFPrrnB-ncl08 colony 11

- iGEMgfp colony 7.1

- iGEMgfp colony 11.1

- iGEMcherry colony 13.1

- iGEMcherry colony 13.2