Latest revision as of 17:24, 21 October 2009

Meetings Calendar

Formal Lab Session - 6th October 2009

Today we did B. subtilis transformations using our ptotocol. We prepared the LB+ CHL(Chloramphenicol) plates since we will transform the cells with pGFP-rrnB. We used 5ml 2.5 mg/ml stock of CHL, however it should have been 2 ml since the final concentration should have been 5ug/ml CHL.

We did miniprep to look for CotC+ pmutin4 colonies. We used 12 ON cultures. Finally we did a restriction digest with HindIII and BamHI. But we could not find a right colony again. Although the transformations seemed worked we had colonies on both "backbone+ no ligase + no insert" and "backbone+ ligase + no insert". Our transformations dd not have the ligated product! Digest:

H20 11ul
Buffer 2ul
DNa 5ul
HindIII 1ul
BamHI 1ul

Control Digest:

H20 13ul
Buffer 2ul
uncut pmutin4 2ul
HindIII 1ul
BamHI 1ul

The bands were pmutin4 were not very clear and we decided to do a midiprep for pmutin4 again.

Experiment procedure

B.subtilis transformation

 1. kinA+pGFP-rrnB from geneArt           10ul    LB+CHL 100ul
 2. pGFP-rrnB plasmid                      5ul
 3. pSB1AT3                                5ul    LB+CHL 200ul
 4. water                                 10ul

 The concentration of CHL was too high for transformed cells to grow.

E.coli transformation

 1. kinA + pGFP-rrnB backbone             20ul    LB+CHL plate
 2. psc fragment + pMutin4 backbone       20ul    LB+Amp plate
 3. kinA+pGFP-rrnB from geneArt            2ul    LB+Amp plate and LB+CHL plate
 4. cotC+pSB1AT3 from geneArt              2ul    LB+Amp plate
 5. pSB1AT3                               10ul    LB+Amp plate
 6. water                                 10ul    LB+Amp plate

July
M	T	W	T	F	S	S
		[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/1_July_2009&action=edit 1]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/2_July_2009&action=edit 2]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/3_July_2009&action=edit 3]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/4_July_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/5_July_2009&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/6_July_2009&action=edit 6]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/7_July_2009&action=edit 7]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/8_July_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/9_July_2009&action=edit 9]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/10_July_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/11_July_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/12_July_2009&action=edit 12]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/13_July_2009&action=edit 13]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/14_July_2009&action=edit 14]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/15_July_2009&action=edit 15]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/16_July_2009&action=edit 16]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/17_July_2009&action=edit 17]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/18_July_2009&action=edit 18]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/19_July_2009&action=edit 19]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/20_July_2009&action=edit 20]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/21_July_2009&action=edit 21]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/22_July_2009&action=edit 22]	[http://2009.igem.org/Team:Newcastle/Labwork/23_July_2009 23]	[http://2009.igem.org/Team:Newcastle/Labwork/24_July_2009 24]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/25_July_2009&action=edit 25]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/26_July_2009&action=edit 26]
[http://2009.igem.org/Team:Newcastle/Labwork/27_July_2009 27]	[http://2009.igem.org/Team:Newcastle/Labwork/28_July_2009 28]	[http://2009.igem.org/Team:Newcastle/Labwork/29_July_2009 29]	[http://2009.igem.org/Team:Newcastle/Labwork/30_July_2009 30]	[http://2009.igem.org/Team:Newcastle/Labwork/31_July_2009 31]

August
M	T	W	T	F	S	S
					[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/1_August_2009&action=edit 1]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/2_August_2009&action=edit 2]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/3_August_2009&action=edit 3]	[http://2009.igem.org/Team:Newcastle/Labwork/4_August_2009 4]	[http://2009.igem.org/Team:Newcastle/Labwork/5_August_2009 5]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/6_August_2009&action=edit 6]	[http://2009.igem.org/Team:Newcastle/Labwork/7_August_2009 7]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/8_August_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/9_August_2009&action=edit 9]
[http://2009.igem.org/Team:Newcastle/Labwork/10_August_2009 10]	[http://2009.igem.org/Team:Newcastle/Labwork/11_August_2009 11]	[http://2009.igem.org/Team:Newcastle/Labwork/12_August_2009 12]	[http://2009.igem.org/Team:Newcastle/Labwork/13_August_2009 13]	[http://2009.igem.org/Team:Newcastle/Labwork/14_August_2009 14]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/15_August_2009&action=edit 15]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/16_August_2009&action=edit 16]
[http://2009.igem.org/Team:Newcastle/Labwork/17_August_2009 17]	[http://2009.igem.org/Team:Newcastle/Labwork/18_August_2009 18]	[http://2009.igem.org/Team:Newcastle/Labwork/19_August_2009 19]	[http://2009.igem.org/Team:Newcastle/Labwork/20_August_2009 20]	[http://2009.igem.org/Team:Newcastle/Labwork/21_August_2009 21]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/22_August_2009&action=edit 22]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/23_August_2009&action=edit 23]
[http://2009.igem.org/Team:Newcastle/Labwork/24_August_2009 24]	[http://2009.igem.org/Team:Newcastle/Labwork/25_August_2009 25]	[http://2009.igem.org/Team:Newcastle/Labwork/26_August_2009 26]	[http://2009.igem.org/Team:Newcastle/Labwork/27_August_2009 27]	[http://2009.igem.org/Team:Newcastle/Labwork/28_August_2009 28]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/29_August_2009&action=edit 29]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/30_August_2009&action=edit 30]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/31_August_2009&action=edit 31]

September
M	T	W	T	F	S	S
	[http://2009.igem.org/Team:Newcastle/Labwork/1_September_2009 1]	[http://2009.igem.org/Team:Newcastle/Labwork/2_September_2009 2]	[http://2009.igem.org/Team:Newcastle/Labwork/3_September_2009 3]	[http://2009.igem.org/Team:Newcastle/Labwork/4_September_2009 4]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/5_September_2009&action=edit 5]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/6_September_2009&action=edit 6]
[http://2009.igem.org/Team:Newcastle/Labwork/7_September_2009 7]	[http://2009.igem.org/Team:Newcastle/Labwork/8_September_2009 8]	[http://2009.igem.org/Team:Newcastle/Labwork/9_September_2009 9]	[http://2009.igem.org/Team:Newcastle/Labwork/10_September_2009 10]	[http://2009.igem.org/Team:Newcastle/Labwork/11_September_2009 11]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/12_September_2009&action=edit 12]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/13_September_2009&action=edit 13]
[http://2009.igem.org/Team:Newcastle/Labwork/14_September_2009 14]	[http://2009.igem.org/Team:Newcastle/Labwork/15_September_2009 15]	[http://2009.igem.org/Team:Newcastle/Labwork/16_September_2009 16]	[http://2009.igem.org/Team:Newcastle/Labwork/17_September_2009 17]	[http://2009.igem.org/Team:Newcastle/Labwork/18_September_2009 18]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/19_September_2009&action=edit 19]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/20_September_2009&action=edit 20]
[http://2009.igem.org/Team:Newcastle/Labwork/21_September_2009 21]	[http://2009.igem.org/Team:Newcastle/Labwork/22_September_2009 22]	[http://2009.igem.org/Team:Newcastle/Labwork/23_September_2009 23]	[http://2009.igem.org/Team:Newcastle/Labwork/24_September_2009 24]	[http://2009.igem.org/Team:Newcastle/Labwork/25_September_2009 25]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/26_September_2009&action=edit 26]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/27_September_2009&action=edit 27]
[http://2009.igem.org/Team:Newcastle/Labwork/28_September_2009 28]	[http://2009.igem.org/Team:Newcastle/Labwork/29_September_2009 29]	[http://2009.igem.org/Team:Newcastle/Labwork/30_September_2009 30]

October
M	T	W	T	F	S	S
			[http://2009.igem.org/Team:Newcastle/Labwork/1_October_2009 1]	[http://2009.igem.org/Team:Newcastle/Labwork/2_October_2009 2]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/3_October_2009&action=edit 3]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/4_October_2009&action=edit 4]
[http://2009.igem.org/Team:Newcastle/Labwork/5_October_2009 5]	[http://2009.igem.org/Team:Newcastle/Labwork/6_October_2009 6]	[http://2009.igem.org/Team:Newcastle/Labwork/7_October_2009 7]	[http://2009.igem.org/Team:Newcastle/Labwork/8_October_2009 8]	[http://2009.igem.org/Team:Newcastle/Labwork/9_October_2009 9]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/10_October_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/11_October_2009&action=edit 11]
[http://2009.igem.org/Team:Newcastle/Labwork/12_October_2009 12]	[http://2009.igem.org/Team:Newcastle/Labwork/13_October_2009 13]	[http://2009.igem.org/Team:Newcastle/Labwork/14_October_2009 14]	[http://2009.igem.org/Team:Newcastle/Labwork/15_October_2009 15]	[http://2009.igem.org/Team:Newcastle/Labwork/16_October_2009 16]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/17_October_2009&action=edit 17]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/18_October_2009&action=edit 18]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/19_October_2009&action=edit 19]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/20_October_2009&action=edit 20]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/21_October_2009&action=edit 21]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/22_October_2009&action=edit 22]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/23_October_2009&action=edit 23]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/24_October_2009&action=edit 24]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/25_October_2009&action=edit 25]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/26_October_2009&action=edit 26]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/27_October_2009&action=edit 27]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/28_October_2009&action=edit 28]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/29_October_2009&action=edit 29]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/30_October_2009&action=edit 30]	[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/31_October_2009&action=edit 31]

20 – 21 June 2009 - Europe workshop (London)
23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
23 October Practice Presentation (Newcastle)
23 October T-shirts are ready
27 October Practice Presentation (Sunderland)
27 October Poster is ready
30 October – 2 November 2009 - Jamboree (Boston)

Newcastle iGEM Twitter
[http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
[http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]

@@ Line 2: / Line 2: @@
 {{:Team:Newcastle/Header}}
 {{:Team:Newcastle/Left}}
+__NOTOC__
 [[Image:Team Newcastle 2009 iGEM ProbationaryP-Sign.PNG|50px|right]]
 =Formal Lab Session - 6th October 2009=
-Today we did ''B. subtilis'' transformations using our ptotocol. We prepared the LB+ CHL(Chloramphenicol) plates since we will transform the cells with pgfp-rrnb.  We used 5ml 2.5 mg/ml stock of CHL, however it should have been 2 ml since the final concentration should have been 5ug/ml CHL.
+Today we did ''B. subtilis'' transformations using our ptotocol. We prepared the LB+ CHL(Chloramphenicol) plates since we will transform the cells with ''pGFP-rrnB''.  We used 5ml 2.5 mg/ml stock of CHL, however it should have been 2 ml since the final concentration should have been 5ug/ml CHL.
-We did miniprep to look for CotC+ pmutin4 colonies. We used 12 ON cultures. Finally we did a restriction digest with HindIII and BamHI. But we could not find a right colony again. Although the transformations seemed worked we had colonies on both "backbnone+ no ligase + no insert" and "backbnone+ ligase + no insert". Our transformations dd not have the ligated product!
+We did miniprep to look for CotC+ pmutin4 colonies. We used 12 ON cultures. Finally we did a restriction digest with HindIII and BamHI. But we could not find a right colony again. Although the transformations seemed worked we had colonies on both "backbone+ no ligase + no insert" and "backbone+ ligase + no insert". Our transformations dd not have the ligated product!
 Digest:
   H20 11ul

Team:Newcastle/Labwork/6 October 2009

From 2009.igem.org

Latest revision as of 17:24, 21 October 2009

Formal Lab Session - 6th October 2009

Experiment procedure

B.subtilis transformation

E.coli transformation