Hi, I'm studying fungal endophytes isolated from frozen leaves, and I am following a protocol that submerges the leaves in a 0.5% bleach solution for two minutes for surface-sterilization. However, these hydrated leaf samples were frozen at -80C, and the leaf epidermal cells may have been lysed. I'm worried that overexposure to bleach will destroy the endophytic DNA, and leaving me nothing to extract, sequence, and identify.
Are there ways to effectively surface-sterilize the leafs without damaging the endophytic DNA? Or can I shorten submergence time without overly compromising the surface sterilization?
Hi there. Maybe you take some less valuable leafs freeze them, bleach them and check integrity of genomic DNA on a simple agarose gel. Maybe your worries are irrelevant.
Cheers
Thank you for your reply. I've already did a test run in a way that largely follows your reply, and the gel electrophoresis results mostly showed smearing. As I'm trying survey the diversity of the fungal operational taxonomic units (OTUs), losing many OTUs will be problematic for my results and discussions.
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