Evidence map›Paper›PMID 30140723›Full record

ArticlePractical laboratory medicine2018

Biotinylated amplicon sequencing: A method for preserving DNA samples of limited quantity.

Karen Cravero, Arielle Medford, Aparna Pallavajjala, Jenna Canzoniero, Natasha Hunter, David Chu, Rory L Cochran, Ian Waters, Eric S Christenson, Kelly Kyker-Snowman and 3 more

Open access · goldAbstract read
In one paragraph

Article in Practical laboratory medicine, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact, top 92% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed, 4 citations in OpenAlex.

  1. Article
  2. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

13 authors at 2 institutions in 1 country.

Karen CraveroThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Arielle MedfordThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Aparna PallavajjalaThe Sidney Kimmel Comprehensive Cancer Center, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Jenna CanzonieroDepartment of Medicine, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Natasha HunterThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
David ChuThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Rory L CochranThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Ian WatersThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Eric S ChristensonThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Kelly Kyker-SnowmanThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Berry ButtonThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Alex J ColeThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Ben Ho ParkThe Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Sidney Kimmel Comprehensive Cancer Center · USJohns Hopkins University · US

Funding

VACCINES--A NEW PARADIGM FOR BREAST CANCERP50CA088843 · NCI · JOHNS HOPKINS UNIVERSITY · PI DAVIDSON, NANCY ELLEN · 2000 to 2010
$27.8M
Training Program in Cellular and Molecular MedicineT32GM008752 · NIGMS · JOHNS HOPKINS UNIVERSITY · PI RAO, RAJINI · 2000 to 2019
$11.7M
Circulating plasma tumor DNA as a biomarker for early stage breast cancerR01CA194024 · NCI · VANDERBILT UNIVERSITY MEDICAL CENTER · PI PARK, BEN HO · 2016 to 2020
$1.9M
Molecular complete response in blood as a predictor for pathologic complete response after neoadjuvant therapy for breast cancerR01CA214494 · NCI · VANDERBILT UNIVERSITY MEDICAL CENTER · PI PARK, BEN HO · 2018 to 2023
$1.1M
NCI NIH HHS P50 CA088843NCI NIH HHS R01 CA194024NCI NIH HHS R01 CA214494NIGMS NIH HHS T32 GM008752
6 · The paper itself

Abstract

backgroundGenomic testing is often limited by the exhaustible nature of human tissue and blood samples. Here we describe biotinylated amplicon sequencing (BAmSeq), a method that allows for the creation of PCR amplicon based next-generation sequencing (NGS) libraries while retaining the original source DNA. DESIGN AND

methodsBiotinylated primers for different loci were designed to create NGS libraries using human genomic DNA from cell lines, plasma, and formalin-fixed paraffin embedded (FFPE) tissues using the BAmSeq protocol. DNA from the original template used for each BAmSeq library was recovered after separation with streptavidin magnetic beads. The recovered DNA was then used for end-point, quantitative and droplet digital PCR (ddPCR) as well as NGS using a cancer gene panel.

resultsRecovered DNA was analyzed and compared to the original DNA after one or two rounds of BAmSeq. Recovered DNA revealed comparable genomic distributions and mutational allelic frequencies when compared to original source DNA. Sufficient quantities of recovered DNA after BAmSeq were obtained, allowing for additional downstream applications.

conclusionsWe demonstrate that BAmSeq allows original DNA template to be recovered with comparable quality and quantity to the source DNA. This recovered DNA is suitable for many downstream applications and may prevent sample exhaustion, especially when DNA quantity or source material is limiting.

Indexed as

BAmSeq, Biotinylated amplicon sequencingcfDNA, Circulating cell-free DNAddPCR, Droplet digital PCRDroplet digital PCR (ddPCR)FFPE, Formalin-fixed paraffin embeddedgDNA, Genomic DNANext generation sequencingNGS, Next generation sequencingpDNA, Plasma DNAPlasma DNAqPCR, Quantitative polymerase chain reactionTargeted amplicon sequencing

Identifiers

PMID30140723
PMCPMC6104457
OpenAlexW2885649464

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.