26/07/13

From 2013.igem.org

(Difference between revisions)

Latest revision as of 14:15, 22 August 2013

Home	Team	Official Team Profile	Project	Parts Submitted to the Registry	Modeling	Notebook	Safety	Attributions

Sample	Plasmid Volume(ul)	DNA concentration (ng/ul)
1.1	40	131.8
1.2	40	77.1
1.3	46	56.8
1.4	39	109
1.5	41	129.1
1.6	32	93.3

measured DNA concentration using nanodrop

Restriction enzyme digest of the DNA samples

- enzymes: XbaI, PstI
- master mix:
  - Buffer - 14ul
  - H2O - 88.2ul
  - XbaI - 1.4ul
  - PstI - 1.4ul
  - Total volume - 105ul
Each sample contained 5ul of DNA and 15ul of the master mix.
The total amount of DNA in each sample was 200ng
To calculate it we divided 200ng by the concentration of each sample
The rest of the volume was made up with water to give a total of 5ul.
The samples were then incubated in a water bath at 37C for an hour

Agorose Gel preparation

- - 20ml of 5xTBE
  - 80ml of water
  - 0.8g of agarose
- Boil for 3 minutes, 1 minute at a time.
- Let it cool and add 5ul of ethidium bromide.
- Pour and let it set for 30 mins.
- Adding dye and marker
- Used orange g loading dye.
  - 2ul to each sample
- Used 1kb ladder, 5ul per 1 marker well.

Run the gel
Gel picture

@@ Line 1: / Line 1: @@
-[[File:iGEM_lim_restr_dig_260713.jpg]]
+<div style="font-family:arial;padding:5px;border-radius:5px;border:5px solid #FF2800;">
-#purification of the limonene biobrick - BBa_24
+{| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center"
-*Plasmid Volume
+!align="center"|[[Team:Leicester|Home]]
-**1.1 - 40ul
+!align="center"|[[Team:Leicester/Team|Team]]
-**1.2 - 40ul
+!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Leicester Official Team Profile]
-**1.3 - 46ul
+!align="center"|[[Team:Leicester/Project|Project]]
-**1.4 - 39ul
+!align="center"|[[Team:Leicester/Parts|Parts Submitted to the Registry]]
-**1.5 - 41ul
+!align="center"|[[Team:Leicester/Modeling|Modeling]]
-**1.2 - 32ul
+!align="center"|[[Team:Leicester/Notebook|Notebook]]
+!align="center"|[[Team:Leicester/Safety|Safety]]
+!align="center"|[[Team:Leicester/Attributions|Attributions]]
+|}
+</div>
+{|border=1
+|Sample||Plasmid Volume(ul)||DNA concentration (ng/ul)
+|-
+|1.1||40||131.8
+|-
+|1.2||40||77.1
+|-
+|1.3||46||56.8
+|-
+|1.4||39||109
+|-
+|1.5||41||129.1
+|-
+|1.6||32||93.3
+|}
 *measured DNA concentration using nanodrop
-**DNA concentration (ng)
-**1.1 - 131.8
-**1.2 - 77.1
-**1.3 - 56.8
-**1.4 - 109
-**1.5 - 129.1
-**1.2 - 93.3
-*Restriction enzyme digest of the DNA samples
+==Restriction enzyme digest of the DNA samples==
 **enzymes: XbaI, PstI
 **master mix:
@@ Line 29: / Line 44: @@
 *To calculate it we divided 200ng by the concentration of each sample
 *The rest of the volume was made up with water to give a total of 5ul.
-*The samples were them incubated in a water bath at 37C by an hour
+*The samples were then incubated in a water bath at 37C for an hour
-*Agorose Gel preparation
+==Agorose Gel preparation==
 ***20ml of 5xTBE
 ***80ml of water
@@ Line 37: / Line 52: @@
 **Boil for 3 minutes, 1 minute at a time.
 **Let it cool and add 5ul of ethidium bromide.
-**Pour and let set for 30 mins.
+**Pour and let it set for 30 mins.
-**Adding dye and and marker
+**Adding dye and marker
 **Used orange g loading dye.
 ***2ul to each sample
@@ Line 44: / Line 59: @@
 *Run the gel
+*Gel picture
+[[File:iGEM_lim_restr_dig_260713.jpg]]