Plant Genomic DNA Extraction
Last week friday, we was given a chance on running an experiment- Plant Genomic DNA extraction.... It is very interesting..... and gain a lot of knowledge and skills
Plant
Acacia mangium had been used on this experiment. The experiment started by doing some cleaning on the Acacia leaf then followed cut it into small pieces.... ( This methods make it easy for disrupting the cell wall of the leaves)
Later, Liquid Nitrogen is added into the mortal in order to freeze the leaves... then the leaves is rapidly grind into a fine powder with a pestle and mortar as the liquids nitrogen boils off (Liquids nitrogen is added to keep the powder from thawing while grinding)
* Note: Liquids Nitrogen is extremely harmful and extra precaution must be taken
Remember to wear double layer of Gloves...
Grinding the leaves into fine powder
Deng La..... It is done....
Look!! Our lecture is demonstrated the technique of grinding the leaves to us...
In order to prevent contamination, all of our experiment is done on the laminar flow hood... Due to laminar flow hood is not enough to provide for all 139 of biotechnology students, so we have to share each others.. Queue up in a line...
This is the sample after CIA solution is added and centrifuge at 13000rpm for 5 minutes to obtain supernatant. The supernatant is carefully transfered into the sterile, new, clean 2.0ml microcentrifuge tube
This is the sample after cold isopropanol is added and centrifuge at 13000 rpm for 2 minutes in order to precipitate nucleic acids. Then after that it is store at -20 degree celcius freezer for at least 30 minutes.
Plenty sample have been prepared.
Look!! One of friend is pipetting the supernatant out from the centrifuge tube into a new tubes...
After 30 minutes, centrifugation at 13000 rpm for 2 minutes to obtain the pellet DNA. After the pellet of DNA is obtained, the supernatant is discarded. Then DNA wash buffer is added and rinse the pellet by tapping the tube gently with finger tips ( the dilute ethanol removes salts ).
After that, centrifuge again at 13000rpm for 2 minutes. Supernatant is discarded and let the pellet air dry before resuspend in TE buffer for further analysis.
It is ice cream that i brought for her... IT CHOCOLATE>>> HER FAVOR
I and her friends are gathering together to celebrate with her....
One of my friend is designing the cover page of our scrapbooks!!
Then, some of the minced liver tissues is transferred into sterile 1.5ml microcentrifuge tube... Lysis Buffer is added....
After incubation for 1 hours, Phenol:Chloroform is added and it must be done in the Laminar flow hood....
Then, centrifuge the sample and u will obtain the results as below....
Since almost 70 % of students think that CAIS located at a strategic place and very convenient for them to come to CAIS but so sad to see that only about 15% of students always visit CAIS....
Wah!!! Almost Half of students in UNIMAS never used discussion room and even personal carrel room...
Liquids Nitrogen Tank
Later, Liquid Nitrogen is added into the mortal in order to freeze the leaves... then the leaves is rapidly grind into a fine powder with a pestle and mortar as the liquids nitrogen boils off (Liquids nitrogen is added to keep the powder from thawing while grinding)
Deng La..... It is done....
Look!! Our lecture is demonstrated the technique of grinding the leaves to us...
This is the sample after CIA solution is added and centrifuge at 13000rpm for 5 minutes to obtain supernatant. The supernatant is carefully transfered into the sterile, new, clean 2.0ml microcentrifuge tube
This is the sample after cold isopropanol is added and centrifuge at 13000 rpm for 2 minutes in order to precipitate nucleic acids. Then after that it is store at -20 degree celcius freezer for at least 30 minutes.
