Hi all,
there's a lot of chat about extraction tekkers to get HMW DNA so i think we've got that covered; however what i can't find is anything on sampling strategies.
Specifically what is the best way to go from raw tissue to HMW DNA if you CAN'T extract immediately after collection and then sequence straight away.
Is it best to freeze tissues (and at what temp, does tissue type [plant/animal etc] affect the outcome) close to collection and store until extraction/sequencing?
Or is it better to sample and extract immediately then store the DNA (4/-20/-80/LN? and what buffer? or plugs?) until sequencing? And should you make the libraries before storage or when ready to sequence?
I've heard anecdotally that -20 freezing of DNA can reduce the length of fragments significantly....
any help/advice appreciated
this was not so much of a problem with illumina seq
there's a lot of chat about extraction tekkers to get HMW DNA so i think we've got that covered; however what i can't find is anything on sampling strategies.
Specifically what is the best way to go from raw tissue to HMW DNA if you CAN'T extract immediately after collection and then sequence straight away.
Is it best to freeze tissues (and at what temp, does tissue type [plant/animal etc] affect the outcome) close to collection and store until extraction/sequencing?
Or is it better to sample and extract immediately then store the DNA (4/-20/-80/LN? and what buffer? or plugs?) until sequencing? And should you make the libraries before storage or when ready to sequence?
I've heard anecdotally that -20 freezing of DNA can reduce the length of fragments significantly....
any help/advice appreciated
this was not so much of a problem with illumina seq
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