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The TE buffer, as placed in the TE well, consisted of 25 mM HEPES, 50 mM Tris, and varying amounts of target and/or contaminating species.
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As a control, the same volume of M9 buffer was placed in adjacent wells.
A UN buffer force was placed in the Sinai Peninsula.
Subsequently, embryos were genotyped as follows: tissue was placed in tail lysis buffer (100 mM Tris Cl, pH 8.0, 5 mM EDTA, 0.2% SDS, 200 mM NaCl) overnight.
Tissue was prepared as described earlier and placed in a 50 m M phosphate buffer (pH 6.0) with 5% hexadecyltrimethyl ammonium bromide (Sigma Chemical).
At sacrifice, spleen tissue was removed and placed in phosphate buffered saline (PBS) buffer on ice.
Lower respiratory portions were selected from induced sputum samples and placed in an RNA lysis buffer (Buffer RLT-Qiagen, Hilden, Germany) as previously described [10].
As soon as the liver is excised, it is placed in a Petri dish filled with suspension buffer.
A small sample (10 µl) of these cells in cryopreservation mix was placed in 100 µl transport/lysis buffer (as above), and shipped to Cambridge at ambient temperature.
For SRA-based mapping, single worms were placed in 5 μl lysis buffer and processed as above.
Mice were euthanized and the diaphragm was dissected with rib attachments and central tendon intact, and placed in K-H buffer at 37°C as previously described.
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