Sterilization of Plant Tissue and Plant Cell Propagation

10/23/2012

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Purpose: 

Sterilize harvested African Violet leaf and implant in cell culture dish for propagation.



Materials: 

Gloves

Forceps

Scalpel

Bunsen Burner

Striker

70% EtOH

1% Bleach Solution

Beaker

Sterile Water

Sterile Petri Dish with nutrient media(cm)

Parafilm



Method:



1. Cut one fully expanded African Violet leaf. Used a healthy section. Strip divided into 8 pieces.



2. Used forceps to dip leaf in 70% EtOH and then transferred to beaker holding 1% Bleach Solution. Forceps were sterilized using Bunsen burner flame prior to use.



3. After 15 minutes in 1% Bleach Solution with periodic swirling I rinsed in sterile water and left leaf there for approximately 1 minute.



4. Used sterile forceps to remove leaf pieces from sterile water and place them in plate media with approximately 1/2 to 1/3 of the piece in the media. 



5. Wrapped cell culture dish in Parafilm like it was Fort Knox.



6. Labeled plate and placed under grow lights in the bio supply room. 



Data and Results: TBD 



Discussion: 



I deviated from the written procedure by cutting the leaf into smaller sections before sterilizing. This may have only opened up additional surfaces for potential contamination. 



I also deviated from the procedure in not soaking the leaf section in 0.5% bleach solution because there was none available.



​Conclusion: TBD