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文献和实验DNA extraction from blister rust aeciospores and urediniospores using dry grindi
all equipment, surfaces, gloves, etc. that spores may have contacted. Resuspend material that has settled in the tubes after all six samples have been processed. 5) Heat treatment: Incubate tubes in 65 ℃ hot block for 1 hr. Flick tubes to mix every 10 - 15 min
The ribonuclease protection assay (RPA)
, again allowing the temperature to ramp down slowly. All incubations may also be carried out in a water bath. RNase Treatments: 14. Prepare the RNase cocktail (per 20 samples) 2.5 ml RNase buffer 6 µl RNase A + T1 mix Remove
Replication timing by comparative hybridization
the culture to the 37o bath. 5. Once the culture has reached 37o C (it takes 10-15 min), add Pronase (0.01-0.05 mg/ml final conc) to release the cells from the alpha factor block. [You can add dry Pronase. I usually dissolve the Pronase in 5 ml
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