Difference: ADP1Tn-seqLibraryPrep (3 vs. 4)

Revision 42021-03-23 - IsaacGifford

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META TOPICPARENT name="ProtocolList"

Generating Illumina Sequencing Libraries for transposons created in A. baylyi ADP1 with the pBT20 vector.

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This protocol prepares Illumina sequencing libraries from Acinetobacter baylyi ADP1 mutagenized with the pBT20 conjugation vector. It is based on the tn-seq approach used by Eli Powell in the Moran lab in Snodgrassella alvi. Note: low retention eppindorf tubes and sterile filter tips should be used to avoid contaminating reagents and cross contaminating samples.
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This protocol prepares Illumina sequencing libraries from Acinetobacter baylyi ADP1 mutagenized with the pBT20 conjugation vector. It is based on the tn-seq approach used by Eli Powell in the Moran lab in Snodgrassella alvi. Note: low retention eppindorf tubes and sterile filter tips should be used to avoid contaminating reagents and cross contaminating samples.
 

Uncommon lab materials needed before starting

  • Covaris tubes for shearing
Line: 106 to 106
 
  • Go to step 2 19X
  • Hold at 16C
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Once complete, transfer the reaction to an eppindorf and remove the beads with a magnet. Clean up with 60 L AMPure beads and elute in 25 L dH2O. Qubit and submit a sample to the core for bioanayzer analysis. This can be done on a pooled library sample.
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Once complete, transfer the reaction to an Eppindorf and remove the beads with a magnet. Clean up with 60 L AMPure beads and elute in 25 L dH2O. Qubit and submit a sample to the core for bioanayzer analysis. This can be done on a pooled library sample.
  -- Main.BrianRenda - 07 Apr 2016
 
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