26/07/13

From 2013.igem.org

(Difference between revisions)

Revision as of 09:38, 29 July 2013

purification of the limonene biobrick - BBa_24

Plasmid Volume

-1.1 - 40ul -1.2 - 40ul -1.3 - 46ul -1.4 - 39ul -1.5 - 41ul -1.2 - 32ul

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

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 them incubated in a water bath at 37C by 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 set for 30 mins.
Adding dye and and marker
Used orange g loading dye.
- 2ul to each sample

Used 1kb ladder, 5ul per 1 marker well.

Run the gel

@@ Line 10: / Line 10: @@
 #measured DNA concentration using nanodrop
 *DNA concentration (ng)
--1.1 - 131.8
+**1.1 - 131.8
--1.2 - 77.1
+**1.2 - 77.1
--1.3 - 56.8
+**1.3 - 56.8
--1.4 - 109
+**1.4 - 109
--1.5 - 129.1
+**1.5 - 129.1
--1.2 - 93.3
+**1.2 - 93.3
 #Restriction enzyme digest of the DNA samples
 *enzymes: XbaI, PstI
 *master mix:
-Buffer - 14ul
+**Buffer - 14ul
-H2O - 88.2ul
+**H2O - 88.2ul
-XbaI - 1.4ul
+**XbaI - 1.4ul
-PstI - 1.4ul
+**PstI - 1.4ul
-Total volume - 105ul
+**Total volume - 105ul
-Each sample contained 5ul of DNA and 15ul of the master mix.
+*Each sample contained 5ul of DNA and 15ul of the master mix.
-The total amount of DNA in each sample was 200ng
+*The total amount of DNA in each sample was 200ng
-To calculate it we divided 200ng by the concentration of each sample
+*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 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 them incubated in a water bath at 37C by an hour
-*Agorose Gel preparation
+#Agorose Gel preparation
--20ml of 5xTBE
+**20ml of 5xTBE
--80ml of water
+**80ml of water
--0.8g of agarose
+**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 set for 30 mins.
+*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.
 *Adding dye and and marker
-Used orange g loading dye.
+*Used orange g loading dye.
--2ul to each sample
+**2ul to each sample
-Used 1kb ladder, 5ul per 1 marker well.
+*Used 1kb ladder, 5ul per 1 marker well.
-*Run the gel
+#Run the gel