The team's work is difficult and very dangerous, as great blocks of ice are extracted from lakes and transported to be sculpted at the Festival's site.
Regions of the resistant determinant/ int-containing contigs that did not show similarity to any part of the R6 genome or PMEN1 ICE were extracted and queried against the Genbank database by BLASTn.
To create ice roads, water is either pumped from lakes or ice aggregate is extracted from lake ice during the winter months.
Then the contours of single ice floes were extracted (Fig. 1) using the Potrace algorithm51.
After boiling for 1 h, the mixture was placed on ice and proline was extracted using 4 mL of toluene.
The reactions were stopped by placing the tubes on ice and steroids were extracted twice with methylene chloride and dried under nitrogen or by using a rotational vacuum concentrator RVC 2 18 (Christ, Germany).
Samples were placed on ice and plasma was extracted within 30 minutes of collection.
Dissected brains were immediately frozen on dry ice, and RNA was extracted as described previously [ 11].
Tissues were snap frozen on dry ice and DNA was extracted using Qiagen DNeasy kits following manufacturer's protocols.
Subsequently, the tubes were chilled on ice, the DNA was extracted with equal volume of chloroform:isoamylalcohol (24:1) and precipitated with isopropanol for 1 h at −20 °C.
