IGEM:Harvard/2010/Team Vector - Revision history

Aaron Deardon: /* 6-15-2010 */

Aaron Deardon — Wed, 07 Jul 2010 17:35:31 GMT

6-15-2010

←Older revision		Revision as of 17:35, 7 July 2010
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	**Ran digests on 1% agarose gel (protocol link)		**Ran digests on 1% agarose gel (protocol link)
	**Digest successful for both reactions		**Digest successful for both reactions
-	**[[Image:~~061510vectordigest2~~.tiff\|100px]]	+	**[[Image:061510vectordigest3.tiff\|100px]]
	**Cut out backbone bands for both reactions		**Cut out backbone bands for both reactions
	*Annealed open series insert oligos (synthesized by Mr. Gene) (protocol link - karmella's)		*Annealed open series insert oligos (synthesized by Mr. Gene) (protocol link - karmella's)

Aaron Deardon: /* 7-1-2010 */

Aaron Deardon — Tue, 06 Jul 2010 13:39:24 GMT

7-1-2010

←Older revision		Revision as of 13:39, 6 July 2010
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	**Plated on LB+Kan(50ug/ml)+Gent(30ug/ml)+Rif(10ug/ml)		**Plated on LB+Kan(50ug/ml)+Gent(30ug/ml)+Rif(10ug/ml)
	*Poured started cultures in 150mL LB + Kan (50ug/ml)		*Poured started cultures in 150mL LB + Kan (50ug/ml)
		+
		+	==7-2-2010==
		+	*Carried out maxipreps of V9-V12
		+	*Created glycerol stocks of V9-V12
		+
		+	==7-6-2010==
		+	*Agrobacterium plates had colonies for E4 and V10 and no growth for the negative controls
		+	*Sequencing V7 and V8 showed they were correct
		+	*Started cultures for maxiprep of V7 and V8

Jacqueline Y. Quinn: /* 7-1-2010 */

Jacqueline Y. Quinn — Thu, 01 Jul 2010 20:22:57 GMT

7-1-2010

←Older revision		Revision as of 20:22, 1 July 2010
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	==7-1-2010==		==7-1-2010==
-	*Set up starter cultures for V9-V12~~, #5~~ for maxipreps	+	*Set up starter cultures for V9-V12 for maxipreps from colonies from the #5 plate of each construct (in LB+Kan 50ug/ml)
	*Agrobacterium transformation (electroporation)		*Agrobacterium transformation (electroporation)
	**Transformed E4 and V10, plus did a control that was shocked without DNA, and a control that was not shocked but had E4 in it.		**Transformed E4 and V10, plus did a control that was shocked without DNA, and a control that was not shocked but had E4 in it.
	**Plated on LB+Kan(50ug/ml)+Gent(30ug/ml)+Rif(10ug/ml)		**Plated on LB+Kan(50ug/ml)+Gent(30ug/ml)+Rif(10ug/ml)
	*Poured started cultures in 150mL LB + Kan (50ug/ml)		*Poured started cultures in 150mL LB + Kan (50ug/ml)

Jacqueline Y. Quinn: /* 6-30-2010 */

Jacqueline Y. Quinn — Thu, 01 Jul 2010 20:21:40 GMT

6-30-2010

←Older revision		Revision as of 20:21, 1 July 2010
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	*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.		*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.
	*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.		*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.
		+
		+
		+	==7-1-2010==
		+	*Set up starter cultures for V9-V12, #5 for maxipreps
		+	*Agrobacterium transformation (electroporation)
		+	**Transformed E4 and V10, plus did a control that was shocked without DNA, and a control that was not shocked but had E4 in it.
		+	**Plated on LB+Kan(50ug/ml)+Gent(30ug/ml)+Rif(10ug/ml)
		+	*Poured started cultures in 150mL LB + Kan (50ug/ml)

Jacqueline Y. Quinn: /* 6-30-2010 */

Jacqueline Y. Quinn — Thu, 01 Jul 2010 15:35:53 GMT

6-30-2010

←Older revision		Revision as of 15:35, 1 July 2010
Line 162:		Line 162:

	==6-30-2010==		==6-30-2010==
-	Got back sequencing results [[Media:Team_vector_sequence1.zip ~~"sequencing"~~]]	+	Got back sequencing results [[Media:Team_vector_sequence1.zip]]
	*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.		*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.
	*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.		*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.

Jacqueline Y. Quinn: /* 6-30-2010 */

Jacqueline Y. Quinn — Thu, 01 Jul 2010 15:35:41 GMT

6-30-2010

←Older revision		Revision as of 15:35, 1 July 2010
Line 162:		Line 162:

	==6-30-2010==		==6-30-2010==
-	Got back sequencing results [[Media:Team_vector_sequence1.zip]]	+	Got back sequencing results [[Media:Team_vector_sequence1.zip "sequencing"]]
	*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.		*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.
	*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.		*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.

Jacqueline Y. Quinn: /* 6-30-2010 */

Jacqueline Y. Quinn — Thu, 01 Jul 2010 15:34:01 GMT

6-30-2010

←Older revision		Revision as of 15:34, 1 July 2010
Line 162:		Line 162:

	==6-30-2010==		==6-30-2010==
-	Got back [[Media:Team_vector_sequence1.~~zip‎ sequencing results~~]]	+	Got back sequencing results [[Media:Team_vector_sequence1.zip]]
	*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.		*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.
	*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.		*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.

Jacqueline Y. Quinn: /* 6-30-2010 */

Jacqueline Y. Quinn — Thu, 01 Jul 2010 15:33:31 GMT

6-30-2010

←Older revision		Revision as of 15:33, 1 July 2010
Line 162:		Line 162:

	==6-30-2010==		==6-30-2010==
-	Got back sequencing results	+	Got back [[Media:Team_vector_sequence1.zip‎ sequencing results]]
	*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.		*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.
	*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.		*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.

Jacqueline Y. Quinn at 14:01, 1 July 2010

Jacqueline Y. Quinn — Thu, 01 Jul 2010 14:01:16 GMT

←Older revision		Revision as of 14:01, 1 July 2010
Line 160:		Line 160:
	*Mixed required quantities and concentrations of each vector and primer in PCR tubes		*Mixed required quantities and concentrations of each vector and primer in PCR tubes
	*Sent tubes to GeneWiz for sequencing		*Sent tubes to GeneWiz for sequencing
		+
		+	==6-30-2010==
		+	Got back sequencing results
		+	*Confirmed V9, V10, V11, V12 - both #5,6 for all. V11 had a single base pair change in the middle of the GusA sequence (bp 6315, T instead of C). This basepair is at the end of a codon, and the change does not change the translation of the sequence.
		+	*V7, V8 had faulty primers - the reverse was not the reverse compliment and the forward was too close to the sequence, so redesigned primers and ordered them.

Aaron Deardon: /* Expression Series */

Aaron Deardon — Tue, 29 Jun 2010 18:16:52 GMT

Expression Series

←Older revision		Revision as of 18:16, 29 June 2010
Line 3:		Line 3:
	==Expression Series==		==Expression Series==
	[[Image:Expression.png\|900px]]		[[Image:Expression.png\|900px]]
		+	==Reporter Series==
		+	[[Image:Reporter.png\|900px]]

	==Abstract==		==Abstract==