Lab Protocols
Permanent link for this collectionhttps://hdl.handle.net/2022/15255
Disclaimer
These protocols are presented so that others may adapt and possibly improve upon them. Every effort was made to make them accurate, but in some cases they were a work in progress and should not be relied on without independent validation.
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We recognize and acknowledge that certain audiences may be offended by research on animals and note that throughout the research we have taken all possible measures to preserve the welfare of the birds and cause no undue harm or interference. We conduct our research under the oversight of a federally mandated animal care committee, and we hope that our findings will ultimately be helpful to the animals themselves by learning more about their natural history and how best to contribute to their conservation.
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CC BY-NC-SA: http://creativecommons.org/licenses/by-nc-sa/3.0/
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Item type: Item , Steroid Label Clean Up Protocol(2013-03-29) Unsure original creator, possibly updated by Kristal Cain?Protocol for cleaning radio-labeled steroid hormoneItem type: Item , Protocol for steroid radioimmunoassay of plasma(2013-03-29) Joe Casto, Jodie Jawor, Danielle WhittakerThis protocol can be used to measure the concentration of steroid hormones in small plasma samples. This protocol has been copied and modified from protocols provided by Dr. John Wingfield at the University of Washington. It has been written for use in the laboratory of Dr. Ellen Ketterson at Indiana University, and therefore the details may require modification if the procedure is performed elsewhere. The general procedure, which takes 4-5 days to complete, is described below. Prepared January 1998 by Joe Casto, modified December 2005 by Jodie Jawor and May 2010 by Danielle Whittaker.Item type: Item , Salt extraction of DNA from whole blood(2013-03-29) From Peter Dunn's lab; unsure if updated by Kettlab membersProtocol for extracting DNA from whole blood stored in lysis buffer. Uses the salt solution extraction buffer.Item type: Item , Sperm Collecting and Counting Protocol(2013-03-29) George Gee, Elaina Tuttle, Tracey KastProtocol for collecting sperm from songbirds, as well as for counting sperm samples. Also includes data sheet.Item type: Item , Spectrometer Protocol(2013-03-29) Lynn SieffermanProtocol for using the spectrometer to analyze color spectrum from feather samples.Item type: Item , Protocol for steroid radioimmunoassay of egg yolks(2013-03-29) Joe LiparThis protocol can be used to measure the concentration of steroid hormones in yolk samples. This protocol has been copied and modified from protocols provided by Dr. John Wingfield at the University of Washington and Dr. Hubert Schwabl at Washington State University. It has been written for use in the laboratory of Dr. Ellen Ketterson at Indiana University, and therefore the details may require modification if the procedure is performed elsewhere. The general procedure, which takes 5 days to complete, is described below. Variations and additional information are provided in the appendices. Prepared October 1997 by Joe Lipar.Item type: Item , Preen Oil Collection Questions(2013-03-29) Danielle WhittakerQuestions about methodology for collecting preen oil samples from dark-eyed juncos.Item type: Item , Short protocol for direct radioimmunoassay of corticosterone (a.k.a. Little B)(2013-03-29) Joe Casto, Jodie JaworThis protocol can be used to measure the concentration of corticosterone in small plasma samples. This protocol has been copied and modified from protocols provided by Dr. John Wingfield at the University of Washington. It has been written for use in the laboratory of Dr. Ellen Ketterson at Indiana University, and therefore the details may require modification if the procedure is performed elsewhere. The general procedure, which takes 4 days to complete, is described below. Variations and additional information are provided in the appendices. Prepared October 1999 by Joe Casto, modified October 2003 by Jodie Jawor.Item type: Item , RNA extraction protocols(2013-03-29) Christine Bergeon Burns, Kimberly RosvallProtocol for extraction of RNA from avian tissue using Trisol and includes instructions for gel electrophoresis.Item type: Item , Junco Female Ethogram(2013-03-29) Ian Parker-RengaAn ethogram describing various categories of observed behavior, specific for female juncos.Item type: Item , Excel worksheet for RIA(2013-03-29) Steve SchoechA spreadsheet for calculating steroid hormone concentrations from radioimmunoassay results. Calculates concentrations of progesterone, testosterone, estradiol, corticosterone, and dihydrotestosterone.Item type: Item , Egg Yolk preparation(2013-03-29) Unsure original creator, possibly updated by Kristal Cain?Protocol to prepare frozen egg yolks for use in a variety of laboratory tests (ELIZA, Hormone assay, Carotenoid HPLC)Item type: Item , Recipe for Longmire's solution(2013-03-29) Ketterson/Nolan research group, Sara SchrockRecipe for making Longmire's solution, a cell lysis buffer for storing blood samples for DNA analysis. Also includes recipes for stock solutions used in making Longmire's solution.Item type: Item , Molecular sexing of birds(2013-03-29) Danielle WhittakerProtocol for PCR of extracted avian DNA using primers that can determine the sex of the samples. Uses agarose gels to determine the amplified alleles.Item type: Item , Microsatellite PCR and ABI3730 Genotyping Instructions(2013-03-29) Danielle Whittaker, Nicole GerlachProtocol for PCR of extracted avian DNA. Includes directions for PCR to amplify microsatellite loci (used for paternity analysis) and for PCR to amplify sex-specific loci (used for determining sample sex). Includes directions for PCR reactions as well as analysis of PCR product on the ABI3730. Also includes information on the primers used by the Ketterson lab for analyzing junco microsatellite loci.Item type: Item , Blood Sample Collection for DNA(2013-03-29) Ketterson/Nolan research groupProtocol for collecting blood samples for later DNA extractionItem type: Item , Protocol for phenol-chloroform extraction of DNA from whole blood samples(2013-03-29) Danielle WhittakerProtocol for extracting DNA from whole blood stored in lysis buffer. Uses the phenol-chloroform method of extraction.Item type: Item , Protocol for phenol-chloroform extraction of DNA from whole blood samples, with checklist(2013-03-29) Nicole GerlachProtocol for extracting DNA from whole blood stored in lysis buffer. Uses the phenol-chloroform method of extraction. Modified from full protocol to include checklist for each step.Item type: Item , qPCR protocol(2013-03-29) Christine Bergeon Burns, Kimberly RosvallProtocol for quantitative PCR to analyse RNA concentration from cDNA.Item type: Item , Protocol for kit extraction of DNA from whole blood samples(2013-03-29) Nicole GerlachProtocol for extracting DNA from whole blood stored in lysis buffer. Uses IBI Scientific MINI Genomic DNA extraction kits. Includes checklist for each step.