Tag Archives: 2SN

DNA Extraction 12/2/15

The four oyster DNA samples ( # 100, 2HL 1 – 3) incubated at 60 degree for an hour. Samples were previously stored at -80 degrees.

DNA was extracted from them using the mollusc DNA extraction kit. None of the optional steps were performed.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling Box #2’.

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DNA Extraction – 11/19/15

The six oyster DNA samples ( # 94 – 99) incubated at 60 degree for an hour. Samples were previously stored at -80 degrees.

DNA was extracted from them using the mollusc DNA extraction kit. None of the optional steps were performed.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling Box #2’.

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DNA Extraction – 11/18/15

The six oyster DNA samples ( # 88 – 93) incubated at 60 degree for an hour. Samples were previously stored at -80 degrees.

DNA was extracted from them using the mollusc DNA extraction kit. None of the optional steps were performed.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling Box #2’.

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DNA extraction- 11/17/15

The six oyster DNA samples ( # 82 – 87) incubated at 60 degree for an hour. Samples were previously stored at -80 degrees.

DNA was extracted from them using the mollusc DNA extraction kit. None of the optional steps were performed.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling Box #2’.

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DNA Extraction 11/13/15

The six oyster DNA samples ( #76 – 81) incubated at 60 degree for an hour. Samples were previously stored at -80 degrees.

We tried some steps to increase the yield:

1) Centrifuge the sample before adding isopropylalcohol to settle down the tissue. Take only the liquid for rest of the protocol – Didn’t work since tissue and liquid did not separate.

2) Add 200 uL ML1 to sample before adding isopropyl and before centrifuge. – Didn’t work very well 

3) Add 200 uL ML1 buffer after adding isopropyl and centrifuge. Centrifuge again – Worked

DNA was extracted from them using the mollusc DNA extraction kit. None of the optional steps were performed.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling Box 1’.

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DNA Extraction 11/10/15

The eight oyster DNA samples ( #68 – 75) incubated at 60 degree. Samples were previously stored at -80 degrees.

DNA was extracted from them using the mollusc DNA extraction kit. None of the optional steps were performed.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling’.

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DNA Extraction and Opening Oysters 10/28/2015

The six oyster DNA samples ( #22-26) incubated yesterday (10/27/2015) were taken out.

DNA was extracted from them using the mollusc DNA extraction kit. All the optional steps were excluded.

Later the samples were stored in refrigerator in box ‘Oly gDNA Oly Reciprocal Transplant Final Sampling’.

Took out 6 oysters from Fidalgo Bay population for DNA extraction.

I collected gill tissue from oysters #27 – 31 for DNA extraction.

Fidalgo Bay 2SN population: Oysters #27-31

Gill tissues of all oyster were collected in MBL 1 buffer and proteinase K for DNA isolation by  mollusc kit.

Placed all the tubes with gill tissues in 37 C for overnight.

Viscera with digestive glands and gonads, etc. were dissected and discarded to prevent decomposition of rest of body by enzymes. Rest of the body of each oyster was stored in 10X RNAlater solution by weight.

Weighed and recorded following for each oyster:

1) Complete organism

2) Gill tissue collected

3) Body parts collected for storage (excluding viscera)

4) Empty shells

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Opening Oysters 10/27/2015

Took out 5 oysters from Fidalgo Bay population for DNA extraction.

I collected gill tissue from oysters #22 – 26 for DNA extraction.

Fidalgo Bay 2SN population: Oysters #22-16

Gill tissues of all oyster were collected in MBL 1 buffer and proteinase K for DNA isolation by  mollusc kit.

Placed all the tubes with gill tissues in 37 C for overnight.

Viscera with digestive glands and gonads, etc. were dissected and discarded to prevent decomposition of rest of body by enzymes. Rest of the body of each oyster was stored in 10X RNAlater solution by weight.

Weighed and recorded following for each oyster:

1) Complete organism

2) Gill tissue collected

3) Body parts collected for storage (excluding viscera)

4) Empty shells

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DNA extraction from oysters

Took 6 oysters from 4SN population collected at Manchester for DNA extraction —> Found all dead. We have weighed and photo documented them for length/width measurements.

Took two oysters randomly from 2SN population collected at Fidalgo bay (and they were both alive.). Weighed and photo documented them for length/width measurements.

I learned how to open oysters, separate the gill tissue for DNA extraction, and putting specific parts in RNAlater. Stored gill tissue for ~ 5 hours in buffer at 60 C.

Later used DNA extraction kit to extract DNA from both samples. Stored in freezer for further process.

Manchester 4SN oysters – All dead (Considered 4 dead, 2 alive before opening)

Fidalgo Bay-2SN Oysters #1-2 taken for DNA extraction

Manchester 4SN- All dead oysters (Considered 2 live and 4 dead before opening)

 

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