By David Markie
In YAC Protocols skilled researchers provide a complete selection of their simply reproducible and confirmed tools for interpreting advanced human, animal, and plant genomes. The step by step protocols disguise all elements of yeast synthetic chromosomes, starting from the development of YAC libraries to their garage, screening, and database administration, and to their use in such expert purposes because the move of YACs to mammalian cells and the isolation of coding sequences from YACs. YAC Protocols permits staff so as to add promising ways tailored from yeast genetics to their own repertoires and to introduce novel strategies rising from the collecting event with YACs as a device for genome research. Its huge notes and history info not just considerably increase the powerful use of the tools, but in addition offer a helpful hands-on source for these wishing to evolve protocols to fit their very own functions.
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Extra info for YAC Protocols
The cross-section of the gel containing the 23-kb Hind111marker and below is removed from under the preparative lanes. 3. 5 V/cm for another 2 h. Remove the outer hHindII1 lanes, stain, and check that the AHind marker had been successfully electrophoresed back into the slot. Cloning of Telomeres 4. 05MNaCl. Stain the remammg gel to reveal any DNA that has been lost from the blocks. This varies from one block to another; the loss of 25% of the DNA at this stage is likely to be an overestrmate. 5.
Slice away and stain the outer 3LHz:ndIIImarkers (the 600 bp fragment should travel about 5 cm). The cross-section of the gel containing the 23-kb Hind111marker and below is removed from under the preparative lanes. 3. 5 V/cm for another 2 h. Remove the outer hHindII1 lanes, stain, and check that the AHind marker had been successfully electrophoresed back into the slot. Cloning of Telomeres 4. 05MNaCl. Stain the remammg gel to reveal any DNA that has been lost from the blocks. This varies from one block to another; the loss of 25% of the DNA at this stage is likely to be an overestrmate.
The more complex PCR pools based on high density row and From Methods In Molecular Biology, Vol 54. YAC Protocols Edlted by. , Totowa, NJ 33 Cole, Collins, 34 and Dunham High Density YAC Filters B 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 72mm ~-108 mm-* Fig. 1. High-density YAC filters. (A) Representation of a hybridization result showing the 4 x 4 array of 1536 clones gridded from 16 microtiter plates onto a single 8 x 12 cm filter. Autoradiographs and filters are labeled in the top lefthand corner, corresponding to position Al of the microtiter plate.