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Dunn School: Notebook 6, Jul 1986-May 1988

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MS. 12364/7

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Covers experiments 458-469, then the numbering of experiments restarts from number 1 on return from a sabbatical in Melbourne in Aug 1987 (see separate Australian sabbatical notebooks 1 and 2 below).

This notebook covers two independent topics:

1. Finishing off the E coli cosmid mapping and relating the work to a published Cell paper (1986) by Kohara et al (Japan) 2. Starting gene therapy work by ex vivo infection of cells and tissues followed by skin grafting in BALB/C mice
  1. Topic 1:
  2. Frequency of recA+ and LacZ+ clones; sheared libraries in cosmids; looking for overlaps of contigs. Probing for rrn (ribosomal RNA) with Chen Chi (scientific visitor from Beijing) helped particularly in probing cosmids with gene-specific oligo pobes
  3. Experiment 1: Gaps in Japanese Kohara phage λ E. coli map (5 Sep 1987) so that we could publish something from our cosmid contig. database. Completing 8 "gaps" e.g. experiment 8, experiment 16. To do this, the team switched to Hind III and Eco RI mapping on 0.4% agarose gels, so they could compare the cosmids with the Kohara λ restriction data.
  4. Experiment 22: Theoretical calculation to get 99% coverage.
  5. Experiment 24: Discussion with Alan Kendal, influenza section leader, CDC Atlanta.
  6. Topic 2:
  7. Experiments 23-39: Initial advice and factor IX retrovirus vector from Inder Verma (Seattle). This was on advice from Ted Friedmann, who stimulated Brownlee to get into gene therapy for factor IX.
  8. Experiment 33: Skin grafting in BALB/C mice.
  9. Experiment 34: PCR reaction (also using samples from George Petersen, on sabbatical with Brownlee from Dunedin, New Zealand) using 2 dry heating blocks and transferring samples by hand from one to another 29 times. George Petersen's samples had amplified.
  10. Experiments 37, 39A: FIX ELISA assays of mouse blood – negative.

Dates

  • Jul 1986-May 1988

Extent

1 volume

Shelfmark

MS. 12364/7

Repository Details

Part of the Bodleian Libraries Repository

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Weston Library
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Oxford OX1 3BG United Kingdom