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Amaranth/Pollen Monitoring

Amaranth/Pollen Monitoring

Having trouble extracting from such large filters in 50 ml tubes

Possible solution 1

Possible Solution 2

Possible Solution 3

DNA Extraction from pollen

Possible Solution 1 Hurdles:

Locate sonicator compatible with 50 ml tubes

Sort out settings used on sonicator

Possible Solution 2 Hurdles:

Might still be too little volume of extraction buffer after cutting up filter

Possible Solution 3 Hurdles:

Designed for fungal spores

Make 50 mM Tris at pH 8, 50 mM EDTA, 3% SDS, 1 M NaCl solution

5M NaCl on hand, 20% SDS, 1M Tris, .5M EDTA:

 

ml For 500 ml of end solution

 

ml For 500 ml of end solution

1 M Tris

25

.5 M EDTA

50

5 M NaCl

100

20% SDS

75

H2O

250

Potential Amaranth primers:

Amaranth Palmeri specific primers:

Paper for below 3 candidates

ITS

A Palmeri Fwd: GAAGGTGACCAAGTTCATGCTCGGGCGTGGATGGCCTAAAAAG

ITS Amaranth nonPalmeri Fwd: GAAGGTCGGAGTCAACGGATTCGGGCGTGGATGGCCTAAAACA

Universal Rev: ACCAATCGCCGCAGCAGC

Marker 2

Palmeri: GAAGGTGACCAAGTTCATGCTAGGAATGAAAAAGTGTTTAGAGT

nonPalm: GAAGGTCGGAGTCAACGGATTAGGAATGAAAAAGTGTTTAGAGG

Universal Rev: CCCTAAACAAAATCTGCCTACA

Marker3

Palmeri: GAAGGTGACCAAGTTCATGCTAAATTAACGTTAGGAAAGCGT

nonPalm: GAAGGTCGGAGTCAACGGATTAAATTAACGTTAGGAAAGCGG

Universal Rev: ACTCCGACTTGATGAGCTTT