Qiagen DNeasy Blood and Tissue Kit protocol (Limnephilidae)

Knowledge graph centered on Qiagen DNeasy Blood and Tissue Kit protocol (Limnephilidae) with 32 nodes and 111 connections. Top connected: Rocky Mountain Biological Laboratory, Hymenoptera, Lepidoptera, Neothremma, Allomyia.

Method synopsis

Extraction of total genomic DNA from individual caddisfly larvae using Qiagen DNeasy Blood and Tissue kit with automated QiaCube processing after manual tissue homogenization.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Identifying PCR primers to facilitate molecular phylogenetics in Caddisflies (Trichoptera) (2015), Zoological Systematics. Implementations in other papers (listed below) may differ.

1. Extract DNA from specimens
   DNA was prepared from individual specimens of each species using the Qiagen DNeasy Blood and Tissue kit. Each specimen was dissected from its case before being ground using a ceramic mortar and pestle in 180μL of tissue lysis buffer ATL. Once homogenized, 20μL of protein kinase K was added and the mixture was incubated for one hour in a 55°C water bath until tissue was lysed.
   Quantities: 180μL tissue lysis buffer ATL, 20μL protein kinase K per specimen, individual specimens from each of 8 speciesDuration: 1 hour incubation at 55°CConditions: 55°C water bath for tissue lysis
   Equipment: Qiagen DNeasy Blood and Tissue kit, ceramic mortar and pestle, tissue lysis buffer ATL, protein kinase K, 55°C water bath
2. Complete DNA extraction protocol
   Samples were loaded into a Qiagen QiaCube extraction robot and processed using the standard animal tissue extraction protocol for total DNA. DNA yield and quality was checked on a NanoDrop 2000 spectrophotometer. Extracted DNA was stored at -20°C.
   Quantities: Total DNA extracted from individual specimensDuration: Standard QiaCube protocol durationConditions: Laboratory conditions, storage at -20°C
   Equipment: Qiagen QiaCube extraction robot, NanoDrop 2000 spectrophotometer

Typical Equipment

• Qiagen DNeasy Blood and Tissue kit
• ceramic mortar and pestle
• Qiagen QiaCube extraction robot
• NanoDrop 2000 spectrophotometer

Output Measurements

• total genomic DNA
• DNA concentration
• DNA quality metrics

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used