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-	==June 21==	+	==June 21st==
		+
		+	*During this week we began doing 6 hour digests at the recommendation of Dr. Salis to cut down on background
		+
		+
	Sent to the sequencing center:		Sent to the sequencing center:
Line 11:		Line 15:
	7)K081007    796		7)K081007    796
-	*We made 1 liter of LB media.	+	*We made 1 liter of LB media.
-		+	==June 22nd==
-	==June 22==	+
	*Flasks Minipreped:		*Flasks Minipreped:
-	-J23108	+	- J23108
-	-J23116	+	- J23116
-	-J37033	+	- J37033
-	-K091107	+	- K091107
-	-C0261	+	- C0261
-	-K274100	+	- K274100
-	-C3RFP	+	- C3RFP
-	-K112808	+	- K112808
-	*All the minipreps were cut with the following enzymes:	+	*All the minipreps were digested with the following enzymes:
-	-J23108 (SP)	+	- J23108 (SP)
-	-J23116 (SP)	+	- J23116 (SP)
-	-J37033 (XP + phosphatase)	+	- J37033 (XP + phosphatase)
-	-K091107(SP)	+	- K091107(SP)
-	-C0261  (XP + phosphatase)	+	- C0261  (XP + phosphatase)
-	-K274100(XP + phosphatase)	+	- K274100(XP + phosphatase)
-	-C3RFP  (EX)	+	- C3RFP  (EX)
-	-K112808(ES + phosphatase)	+	- K112808(ES + phosphatase)
-	*Ligations:
-	- J23108 + J37033
-	- J23116 + J37033
-	- K091107 + C0261
-	- K091107 + K274100
-	- C3RFP + K112808
-	*Transformations:	+	*We also did a PCR on
-	- B0010    Plate 1 Well 13D	+
-	- R0010    Plate 1 Well 1D	+
-	- K081007  Plate 2 Well 10J	+
-	- J24679  Plate 1 Well 7L	+
		+	-1. K081007 + R0010
		+	-2. J23116 + J37033
		+	-3. J23108 + J37033
		+	<center>[[Image:6-22-10.jpg]]</center>
-	*We also did a PCR on K081007 + R0010, J23116 + J37033, and J23108 + J37033.
-	<center>[[Image:6-22-10.jpg]]</center>	+	*3b and 3c did not turn out for us as planned.
		+	*Ligations:
		+	- J23108 + J37033
		+	- J23116 + J37033
		+	- K091107 + C0261
		+	- K091107 + K274100
		+	- C3RFP + K112808
		+
		+
		+	*Transformations:
		+	- B0010    Plate 1 Well 13D
		+	- R0010    Plate 1 Well 1D
		+	- K081007  Plate 2 Well 10J
		+	- J24679  Plate 1 Well 7L
	* Gel order:		* Gel order:
-	- Ladder	+	- Ladder
-	- J23116	+	- J23116
-	- J23108	+	- J23108
-	- K091107	+	- K091107
-	- C3RFP	+	- C3RFP
		+
		+
	<center>[[Image:6-22-10 001.jpg]]</center>		<center>[[Image:6-22-10 001.jpg]]</center>
-	==June 23==	+	*This gel turned out well for us, C3RFP was visible but faint.
-	*Transform today:	+	==June 23rd==
-	- R0061  Plate 1 Well 14P	+
-	- R0062  Plate 1 Well 6O	+
-	- R0063  Plate 1 Well 8A	+	*Flasks mini prepped:
-	- K091146 Plate 2 Well 8H	+	- K091107
		+	- R0010
		+	- J24679
		+	- J23108
		+	- J23116
		+	- C0261
		+	- K081107
		+	- B0010
		+	- J37033
		+	- K274100
	*Digests:		*Digests:
-	- K091107 (SP)	+	- K091107 (SP)
-	- R0010  (SP)	+	- R0010  (SP)
-	- J24679  (ES)	+	- J24679  (ES)
-	- J23108  (SP)	+	- J23108  (SP)
-	- J23116  (SP)	+	- J23116  (SP)
-	- CO261  (XP)	+	- CO261  (XP)
-	- K081107 (XP)	+	- K081107 (XP)
-	- B0010  (EX)	+	- B0010  (EX)
-	- J37033  (XP)	+	- J37033  (XP)
-	- K274100 (XP)	+	- K274100 (XP)
-	*Sent to get sequenced:	+	*Sent to sequencing center:
-	1) R0010	+	1) R0010
-	2) B0010	+	2) B0010
-	3) K081107	+	3) K081107
-	4) J24679	+	4) J24679
-	==June 24==	+	*Transform today:
		+	- R0061  Plate 1 Well 14P
		+	- R0062  Plate 1 Well 6O
		+	- R0063  Plate 1 Well 8A
		+	- K091146 Plate 2 Well 8H
		+
		+	==June 24th==
	Gel Order:		Gel Order:
-	1) Ladder	+	- Ladder
-	2) J24679	+	- J24679
-	3) K091107	+	- K091107
-	4) J37033	+	- J37033
-	5) J23108	+	- J23108
-	6) K274100	+	- K274100
-	7) B0010	+	- B0010
-	8) R0010	+	- R0010
-	9) K081107	+	- K081107
-	10)CO261	+	- CO261
-	11)J23116	+	- J23116
Line 116:		Line 142:
	RE-DIGEST:		RE-DIGEST:
-	- CO261	+	- CO261
-	- K081107	+	- K081107
	RE-Transform:		RE-Transform:
-	- J37033	+	- J37033
-	- K091107	+	- K091107
	*Sent to sequencing:		*Sent to sequencing:
-	1) R0061	+	1) R0061
-	2) R0062	+	2) R0062
-	3) R0063	+	3) R0063
-	4) K091146	+	4) K091146
-	==June 25==	+	==June 25th==
	*Sent to sequencing:		*Sent to sequencing:
-	1) K091107	+	1) K091107
-	2) J37033	+	2) J37033
	*Digests:		*Digests:
-	- K091107 (SP)	+	- K091107 (SP)
-	- J23108  (SP)	+	- J23108  (SP)
-	- J23116  (SP)	+	- J23116  (SP)
-	- CO261  (XP)	+	- CO261  (XP)
-	- R0010  (SP)	+	- R0010  (SP)
-	- K274100 (XP)	+	- K274100 (XP)
-	- J37033  (XP)	+	- J37033  (XP)
-	- J24679  (ES)	+	- J24679  (ES)
-	- K081007 (XP)	+	- K081007 (XP)
		+
-	*Ligations:
-	- K091107 + K274100
-	- J23108  + J37033
-	- J23116  + J37033
-	- C0261  + J24679
-	- R0010  + K081007
	The 4 that were sent to get sequenced yesterday were ran on a gel. The gel order is provided below:		The 4 that were sent to get sequenced yesterday were ran on a gel. The gel order is provided below:
-	- Ladder	+	- Ladder
-	- K091146	+	- K091146
-	- R0061	+	- R0061
-	- R0062	+	- R0062
-	- R0063	+	- R0063
		+
		+
	<center>[[Image:6-25-10.jpg]]</center>		<center>[[Image:6-25-10.jpg]]</center>
		+
		+
		+
		+
		+	*This gel was perfect for us. We were able to see what we needed to see and extract the correct pieces.
		+
		+	*Ligations:
		+	- K091107 + K274100
		+	- J23108  + J37033
		+	- J23116  + J37033
		+	- C0261  + J24679
		+	- R0010  + K081007
		+

---

Latest revision as of 15:41, 5 August 2010

Contents 1 June 21st 2 June 22nd 3 June 23rd 4 June 24th 5 June 25th

June 21st

• During this week we began doing 6 hour digests at the recommendation of Dr. Salis to cut down on background

Sent to the sequencing center:

              Base Pairs
 1)C3RFP      681
 2)K091146    1266
 3)K112022    1499
 4)K145150    66
 5)R0063      151
 6)R1062      56
 7)K081007    796

• We made 1 liter of LB media.

June 22nd

• Flasks Minipreped:

   - J23108
   - J23116
   - J37033
   - K091107
   - C0261
   - K274100
   - C3RFP
   - K112808

• All the minipreps were digested with the following enzymes:

   - J23108 (SP)
   - J23116 (SP)
   - J37033 (XP + phosphatase)
   - K091107(SP)
   - C0261  (XP + phosphatase)
   - K274100(XP + phosphatase)
   - C3RFP  (EX)
   - K112808(ES + phosphatase)

• We also did a PCR on

   -1. K081007 + R0010 
   -2. J23116 + J37033
   -3. J23108 + J37033

• 3b and 3c did not turn out for us as planned.

• Ligations:

   - J23108 + J37033
   - J23116 + J37033
   - K091107 + C0261
   - K091107 + K274100
   - C3RFP + K112808

• Transformations:

   - B0010    Plate 1 Well 13D
   - R0010    Plate 1 Well 1D
   - K081007  Plate 2 Well 10J
   - J24679   Plate 1 Well 7L

• Gel order:

   - Ladder
   - J23116
   - J23108
   - K091107
   - C3RFP

• This gel turned out well for us, C3RFP was visible but faint.

June 23rd

• Flasks mini prepped:

   - K091107
   - R0010
   - J24679
   - J23108
   - J23116
   - C0261
   - K081107
   - B0010
   - J37033
   - K274100

• Digests:

   - K091107 (SP)
   - R0010   (SP)
   - J24679  (ES)
   - J23108  (SP)
   - J23116  (SP)
   - CO261   (XP)
   - K081107 (XP)
   - B0010   (EX)
   - J37033  (XP)
   - K274100 (XP)

• Sent to sequencing center:

   1) R0010
   2) B0010
   3) K081107
   4) J24679

• Transform today:

   - R0061  Plate 1 Well 14P
   - R0062  Plate 1 Well 6O
   - R0063  Plate 1 Well 8A
   - K091146 Plate 2 Well 8H

June 24th

Gel Order:

   - Ladder
   - J24679
   - K091107
   - J37033
   - J23108
   - K274100
   - B0010
   - R0010
   - K081107
   - CO261
   - J23116

RE-DIGEST:

   - CO261
   - K081107

RE-Transform:

   - J37033
   - K091107

• Sent to sequencing:

   1) R0061
   2) R0062
   3) R0063
   4) K091146

June 25th

• Sent to sequencing:

   1) K091107
   2) J37033

• Digests:

   - K091107 (SP)
   - J23108  (SP)
   - J23116  (SP)
   - CO261   (XP)
   - R0010   (SP)
   - K274100 (XP)
   - J37033  (XP)
   - J24679  (ES)
   - K081007 (XP)

The 4 that were sent to get sequenced yesterday were ran on a gel. The gel order is provided below:

   - Ladder
   - K091146
   - R0061
   - R0062
   - R0063

• This gel was perfect for us. We were able to see what we needed to see and extract the correct pieces.

• Ligations:

   - K091107 + K274100
   - J23108  + J37033
   - J23116  + J37033
   - C0261   + J24679 
   - R0010   + K081007

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