Jones Lab:Protocols:Sterilise seed

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(New page: <H3>Sterilise Seed</H3> 1) 5 min 70% ethanol 2) 10 min 100% ethanol 3) Water washes 4) Stratify 3 days 4°C 5) Plate seed)
Current revision (11:23, 7 October 2013) (view source)
 
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<H3>Sterilise Seed</H3>
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<H3>Sterilise Seed with chlorine gas</H3>
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1) Place small number of seed (max 300/~5mm in eppendorf) and open lids
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2) Place to desiccator
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3) Add 3ml HCl (5M) to 50ml commercial bleach and quickly add desiccator lid
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<font color="red">THIS STEP IN FUME HOOD- CHLORINE GAS PRODUCED</font>
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4) After 3hrs, transfer to sterile hood
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5) Leave for 20mins for residual gas to evaporate, then add ~500ul dH2O
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6) Dispose of HCl/bleach mixture in sink in fume hood WITH EXCESS WATER (~5min of tap running)
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7) Move seed to fridge/cold room for 3 days
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8) Plate
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<H3>Sterilise Seed with Ethanol</H3>
1) 5 min 70% ethanol
1) 5 min 70% ethanol

Current revision

Sterilise Seed with chlorine gas

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

Sterilise Seed with Ethanol

1) 5 min 70% ethanol

2) 10 min 100% ethanol

3) Water washes

4) Stratify 3 days 4°C

5) Plate seed

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