Details:

Alternative Title:
Biopreserv Biobank
Personal Author:
Miernyk, KM ; DeByle, CD ; Rudolph, KM
Miernyk, KM ; DeByle, CD ; Rudolph, KM Less -
Description:
Background:

Culture-independent molecular analyses allow researchers to identify diverse microorganisms This approach requires microbiological DNA repositories The standard for DNA storage is liquid nitrogen or ultra-low freezers These use large amounts of space, are costly to operate, and could fail Room temperature DNA storage is a viable alternative In this study we investigated storage of bacterial DNA using two ambient storage matrices, Biomatrica DNAstable® Plus and GenTegra® DNA

Methods:

We created crude and clean DNA extracts from five Streptococcus pneumoniae isolates Extracts were stored at −30°C (our usual DNA storage temperature), 25°C (within the range of temperatures recommended for the products), and 50°C (to simulate longer storage time) Samples were stored at −30°C with no product and dried at 25°C and 50°C with no product, Biomatrica DNAstable® Plus, or GenTegra® DNA We analyzed the samples after 0, 1, 2, 4, 8, 16, 32, and 64 weeks using the Nanodrop 1000 to determine the amount of DNA in each aliquot and by qPCR for the S pneumoniae genes lytA and psaA Using a 50°C storage temperature, we simulated 362 weeks of 25°C storage

Results:

The average amount of DNA in aliquots stored with a stabilizing matrix was 103%-116% of the original amount added to the tubes This is similar to samples stored at −30°C (average 102%-121%) With one exception, samples stored with a stabilizing matrix had no change in lytA or psaA Ct value over time (Ct range ≤ 29), similar to samples stored at −30°C (Ct range ≤ 30) Samples stored at 25°C with no stabilizing matrix had Ct ranges of 22 – 51

Conclusion:

DNAstable® Plus and GenTegra® DNA can protect dried bacterial DNA samples stored at room temperature with similar effectiveness as at −30°C It is not effective to store bacterial DNA at room temperature without a stabilizing matrix


Subjects:
[+]
Adhesins, Bacterial Article Desiccation DNA, Bacterial Lipoproteins Reagent Kits, Diagnostic Specimen Handling Streptococcus Pneumoniae Temperature
Source:
Biopreserv Biobank 15(6):529-534
Pubmed ID:
29130748
Pubmed Central ID:
PMC6677143
Document Type:
Journal Article
Funding:
CC999999/Intramural CDC HHS/Intramural CDC HHS/United States
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:a4dcad9670266fd83ee0aa4fcdd26bfc7678b6e0e48635263a51db603eaf3dc3
Download URL:
https://stacks.cdc.gov/view/cdc/80406/cdc_80406_DS1.pdf
File Type:

Supporting Files

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