Library Preparation Troubleshooting
Library preparation will likely define the quality of your sequencing data. While the construction of a genomic library will be slightly different from a cDNA library, with different kits for each, some basic principles apply to both. Despite this, things can still go awry.
If this is the case for you, have no fear! This ZAGENO troubleshooting guide is here to help.
Poor DNA quality: Make sure that your material is not degraded and has been stored correctly
Insufficient purity: Be sure to purify your DNA preparation before using it for library creation
Wrong quantification: The quantification of your DNA might not have been executed precisely enough. Check the quality of your quantification
Insufficient amplification: Add two or three amplification steps in your protocol
Insufficient adapter ligation Make sure that your ligation followed the supplier protocol
Too may amplification cycles: Excessive amplification can amplify biased sequence coverage. Reduce the amplification steps in your protocol
Amplification range over instrumental sensitivity: The amplification might have pushed some sequences over the instrumental sensitivity of your instrument. Reduce the amplification steps in your protocol.