Daniel:Protocols/LatchPadlock: Difference between revisions

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>Djacobse
>Djacobse
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<li>After washing, incubate with 100 nM insert/backbone oligos for 30 min at 37C</li>
<li>After washing, incubate with 100 nM insert/backbone oligos for 30 min at 37C</li>
<li>Wash twice</li>
<li>Wash twice</li>
<ol type="A"><li>Wash by pelleting cells at 600xg for 3 min</li></ol>


</ol>
</ol>

Revision as of 23:44, 8 June 2016

Latch and Padlock Hybridization

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Protocol

  1. Start from hybridized C Probes or antibodies
    1. Make sure probes have been washed as per the protocol
  2. After washing, incubate with 100 nM insert/backbone oligos for 30 min at 37C
  3. Wash twice
    1. Wash by pelleting cells at 600xg for 3 min


Buffers

  1. Insert/backbone Hybridization Buffer
    1. 100 nM insert/backbone oligos
    2. 1X SSC
    3. 40 U/mL RNasin
    4. PBS
  2. Wash Buffer
    1. PBS
    2. 0.1% Tween
    3. 4 U/mL RNasin