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: @@
-*this
+<div style="font-family:arial;padding:5px;border-radius:5px;border:5px solid #FF2800;">
-*is
+{| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center"
-*a
+!align="center"|[[Team:Leicester|Home]]
-*numbered
+!align="center"|[[Team:Leicester/Team|Team]]
-*list
+!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Leicester Official Team Profile]
+!align="center"|[[Team:Leicester/Project|Project]]
+!align="center"|[[Team:Leicester/Parts|Parts Submitted to the Registry]]
+!align="center"|[[Team:Leicester/Modeling|Modeling]]
+!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
+==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
+[[File:iGEM_lim_restr_dig_260713.jpg]]