Tag Archives: T3B

DNase Treatment – Water Filter RNA from 20161130

Continued preparation of this RNA to assess withering syndrome viability in the water column. I treated the RNA I isolated on 20161130 using the Turbo DNA-free (Ambion) DNase kit, according to their protocol.

Added the following to each sample:

  • 2.5μL 10x buffer
  • 1.5μL H2O
  • 1μL DNase

Incubated @ 37C for 1hr.

Added 0.1 volumes (2.5μL) of DNase Inactivation reagent and incubated at RT for 2mins (with mixing). Pelleted inactivation reagent: 10,000g, 2mins, RT. Transferred supe to new tube.

Samples were labelled as “DNased RNA”, their existing sample name (see below), and stored @ -80C.

Sample names:

  • T0A
  • T0B
  • T1A
  • T1B
  • T1A C
  • T1B C
  • T3A
  • T3B
  • T7A
  • T7B
  • T7A C
  • T7B C
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RNA Isolation – Abalone Water Filters

Lisa provided me with a set of water filters stored in 1mL RNAzol RT @ -80C to isolate RNA from. This is an attempt to assess withering syndrome viability from within the water.

The samples were thawed and briefly homogenized (as best I could) with a disposable plastic pestle. The samples were then processed according to the manufacturer’s protocol for total RNA isolation. Samples were resuspended in 20μL of 0.1%-DEPC H2O.

Samples were stored @ -80C. Will DNase next week.

The sample names are as follows (the ‘C’ is short for “Control”, per Lisa):

  • T0A
  • T0B
  • T1A
  • T1A C
  • T1B C
  • T3A
  • T3B
  • T7A
  • T7B
  • T7A C
  • T7B C

 

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