Jones Lab:Protocols:Sterilise seed: Difference between revisions

Latest revision as of 09:23, 7 October 2013

1) Place small number of seed (max 300/~5mm in eppendorf) and open lids

2) Place to desiccator

3) Add 3ml HCl (5M) to 50ml commercial bleach and quickly add desiccator lid THIS STEP IN FUME HOOD- CHLORINE GAS PRODUCED

4) After 3hrs, transfer to sterile hood

5) Leave for 20mins for residual gas to evaporate, then add ~500ul dH2O

6) Dispose of HCl/bleach mixture in sink in fume hood WITH EXCESS WATER (~5min of tap running)

7) Move seed to fridge/cold room for 3 days

8) Plate

1) 5 min 70% ethanol

2) 10 min 100% ethanol

3) Water washes

4) Stratify 3 days 4°C

5) Plate seed

@@ Line 1: / Line 1: @@
-<H3>Sterilise Seed</H3>
+<H3>Sterilise Seed with chlorine gas</H3>
+) Place small number of seed (max 300/~5mm in eppendorf) and open lids
+) Place to desiccator
+) Add 3ml HCl (5M) to 50ml commercial bleach and quickly add desiccator lid
+	<font color="red">THIS STEP IN FUME HOOD- CHLORINE GAS PRODUCED</font>
+) After 3hrs, transfer to sterile hood
+) Leave for 20mins for residual gas to evaporate, then add ~500ul dH2O
+) Dispose of HCl/bleach mixture in sink in fume hood WITH EXCESS WATER (~5min of tap running)
+) Move seed to fridge/cold room for 3 days
+) Plate
+<H3>Sterilise Seed with Ethanol</H3>
 ) 5 min 70% ethanol