Newcastle University Wetlab/18 September 2008

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(New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Thursday 18th September== 300px * Plasmid was isolated from 2mL of each of the 6 ON cultures. pGF...)
 
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* Plasmid was [[isolated]] from 2mL of each of the 6 ON cultures. pGFPrrnB-ncl08 colonies 7 + 11 were [[restricted]] using 7.5μl plasmid and 0.5μl each enzyme (EcoRI and NheI) in 15μl total volume.
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* Plasmid was [[Isolating Plasmid from Cells (Miniprep)|isolated]] from 2mL of each of the 6 ON cultures. pGFPrrnB-ncl08 colonies 7 + 11 were [[Restricting Plasmids (Double Restriction)|restricted]] using 7.5μl plasmid and 0.5μl each enzyme (EcoRI and NheI) in 15μl total volume.
Lane 1: 1 kb ladder
Lane 1: 1 kb ladder
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This showed that glycerol stocks of these plasmids are good as the fragment sizes are as expected. pGFPrrnBrrnB colony 7 is particularly strong and contains a large quantity of DNA.
This showed that glycerol stocks of these plasmids are good as the fragment sizes are as expected. pGFPrrnBrrnB colony 7 is particularly strong and contains a large quantity of DNA.
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* iGEMgfp colony 7.1 was [[induced with subtilin]] and [[prepared for microscopy]]. This showed no difference between the subtilin-induced and the control cultures. Also, only some cells expressed ''gfp'' whilst others did not. However, it was noticed that every cell that did flouresce was also undergoing sporulation, possibly indicating that the insert promotor may be under the control of the sporulation factor.
+
* iGEMgfp colony 7.1 was [[Inducing Bacillus subtilis with Subtilin|induced with subtilin]] and [[Preparing Bacillus subtilis for Microscopy|prepared for microscopy]]. This showed no difference between the subtilin-induced and the control cultures. Also, only some cells expressed ''gfp'' whilst others did not. However, it was noticed that every cell that did flouresce was also undergoing sporulation, possibly indicating that the insert promotor may be under the control of the sporulation factor.
* ON agar plates of iGEMBB107 colony 3 and 5 (plated on starch + chloramphenicol, spectinomycin and chloramphenicol) were analysed. Colonies were wanted that did not have a 'halo' on starch + cam, that did not grow on spec and that did grow on cam. All but colony 3.9 showed to be good. Therefore ON cultures were made from the cam only plates of colonies 3.1, 3.2, 5.1 and 5.2 in 3mL LB.
* ON agar plates of iGEMBB107 colony 3 and 5 (plated on starch + chloramphenicol, spectinomycin and chloramphenicol) were analysed. Colonies were wanted that did not have a 'halo' on starch + cam, that did not grow on spec and that did grow on cam. All but colony 3.9 showed to be good. Therefore ON cultures were made from the cam only plates of colonies 3.1, 3.2, 5.1 and 5.2 in 3mL LB.
* ON culture was also made from iGEMcherry colony 13.1 glycerol stock in 3mL LB for microscopy.
* ON culture was also made from iGEMcherry colony 13.1 glycerol stock in 3mL LB for microscopy.

Latest revision as of 16:46, 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]

Thursday 18th September

1809.jpg
  • Plasmid was isolated from 2mL of each of the 6 ON cultures. pGFPrrnB-ncl08 colonies 7 + 11 were restricted using 7.5μl plasmid and 0.5μl each enzyme (EcoRI and NheI) in 15μl total volume.

Lane 1: 1 kb ladder Lane 2: pGFPrrnB-ncl08 colony 7 restricted with EcoRI + NheI Lane 3: pGFPrrnB-ncl08 colony 11 restricted with EcoRI + NheI Lane 4: -

This showed that glycerol stocks of these plasmids are good as the fragment sizes are as expected. pGFPrrnBrrnB colony 7 is particularly strong and contains a large quantity of DNA.

  • iGEMgfp colony 7.1 was induced with subtilin and prepared for microscopy. This showed no difference between the subtilin-induced and the control cultures. Also, only some cells expressed gfp whilst others did not. However, it was noticed that every cell that did flouresce was also undergoing sporulation, possibly indicating that the insert promotor may be under the control of the sporulation factor.
  • ON agar plates of iGEMBB107 colony 3 and 5 (plated on starch + chloramphenicol, spectinomycin and chloramphenicol) were analysed. Colonies were wanted that did not have a 'halo' on starch + cam, that did not grow on spec and that did grow on cam. All but colony 3.9 showed to be good. Therefore ON cultures were made from the cam only plates of colonies 3.1, 3.2, 5.1 and 5.2 in 3mL LB.
  • ON culture was also made from iGEMcherry colony 13.1 glycerol stock in 3mL LB for microscopy.
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