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. Author manuscript; available in PMC: 2009 Jun 30.
Published in final edited form as: Nat Struct Mol Biol. 2007 Dec 23;15(1):34–42. doi: 10.1038/nsmb1354

Figure 1.

Figure 1

Loss of viability and telomeric DNA in cells lacking the candidate gene for telomerase RNA. (a) Two successive immunoprecipitations greatly enrich candidate telomerase RNA (CTR). Top, multiplex RT-PCR analysis of aliquots from input (0.3% of cleared extract), anti-Myc immunoprecipitation (3% of immunoprecipitate) and anti-TMG immunoprecipitation (64% of immunoprecipitate) with oligonucleotides that amplify TER1 and 7SL RNA recovered from a Trt1p-9Myc overexpression strain. Eighty percent of precipitate recovered from the anti-Myc immunoprecipitation was used in the anti-TMG immunoprecipitation. Input and anti-Myc immunoprecipitation aliquots were amplified with 30 cycles and the anti-TMG immunoprecipitation aliquots were amplified with 40 cycles. Bottom, western blot analysis of input (1% of cleared extract) and anti-Myc immunoprecipitation (4% of immunoprecipitate) from a Trt1p-9Myc overexpression strain. *, cross-reacting nonspecific band. (b) Six successive restreaks of the parental WT haploid and an isogenic strain in which the CTR gene was replaced with his3+ shown from top to bottom. Streaks were grown on YES media at 30 °C for 3–4 days. (c) Telomere length analysis of successive streaks of parental WT strain (left) and CTR disrupted strain (right) after successive restreaks. Bracket, position of WT telomere signal. The bands at 2.0 and 5.0 Kb probably result from cross-reaction with the telomeric probes21.