from Dave Matthews, 12/93 revised 4/96 How to Create Probe Records for GrainGenes Here is a description of the fields available for Probe data in GrainGenes. TEMPLATE Probe : "" Other_name "" Locus "" Reference "" General_remark "" Type "" PCR_primers "" Amplification_conditions "" Size Insert_enzyme "" Source_gene_class "" Source_species "" Source_germplasm "" Source_tissue "" Vector "" Vector_enzyme "" Excision_enzyme "" Vector_PCR_primers "" Vector_amplification "" Bacterial_strain "" Antibiotic "" Location "" Authority "" Image "" Data_source "" Date "" Note: this template does not include all of the fields available. The current list is described in EXPLANATION below, and more will probably be added. Fields that have no data should be deleted. EXAMPLE SYNTAX NOTES Probe : "ABC169" First line must include the " : ". Locus "abc169A" If a field has multiple values, list Locus "abc169B" them one after the other like this. Reference "TAG-86-705" General_remark "Steptoe malt cDNA" Type "cDNA" Size 0.3 Do not use quotes for numeric values. Source_species "Hordeum vulgare" Source_germplasm "Steptoe" Source_tissue Malt If the value of a field does not Vector pBS contain any blank spaces, the quotes Excision_enzyme "EcoRI" (") are optional. Location "Kleinhofs, Andris" Data_source "ATMI" Date "92.09" EXPLANATION Column heading Sample Entry Description or List of Choices ----------------------------------------------------------------------------- Probe PSR177 Name of Probe Other_name pDEM47x.2a Any other name by which this Probe is known, esp. if published Locus Xpsr177 Name of Locus. List all loci mapped with the probe. External_DB Sequence L43928 Genbank accession number Reference GNM-34-362 Reference ID for the reference (see "Constructing reference IDs"). Include entire reference as a separate record. General_remark This probe works Any text only on odd days. Type cDNA cDNA, genomic, PCR PCR_primers AATGCCGAAT Sequence of pcr primers (for the probe, not the vector) AFLP_primers E37 Sequence or code for AFLP primers Amplification_conditions Anneal at 50oC,... Description of conditions Specificity Adh1 or 3, not 2 Which genes are known to be recognized or not recognized Sequence X123429 GENBANK accession #, or other name of a Sequence Size 1.2 Sequence length, in kilobases PCR_size 1.4 Length between Vector_PCR_primers Background 3/5 Background hybridization level and scale Cross_hybridizes_to Zea mays Name of a species Insert_enzyme EcoRI Name of enzyme used to cut the source DNA for insertion into the vector (Should we change the name "Insert_enzyme" to something more descriptive? Source_excision_enzyme"? "Library_enzyme"? "Cloning_enzyme"?) DNA_origin HTF-genomic lib. Library preparation method Source_gene Adh1 Name of Gene from which the probe was derived, if known Source_gene_class Adh Name of Gene_Class from which the probe was derived, if actual gene is not known Source_species Triticum aestivum Name of Species source of DNA. There should ALWAYS be a value for this field. Source_germplasm Chinese Spring Name of Germplasm source of DNA Source_tissue etiolated leaves Tissue source of DNA Vector pUC18 Vector in which probe is cloned Vector_enzyme EcoRI Name of enzyme used to open the vector Excision_enzyme EcoRI Name of enzyme used to remove the insert from vector Vector_PCR_primers M13F Names of Sequences in vector to M13R amplify the probe from Vector_amplification 50uM dATP dCTP... Conditions for amplifying from the vector Bacterial_strain XL1 Blue Host for the clone Antibiotic Ampicillin Resistance carried by the vector Location Gill, Bikram S. Name of Colleague from whom the probe can be obtained Authority Gale, Michael D. Name of Colleague who created the probe Subclone_of pPSR77 Name of original Probe from which PSR177 was subcloned Subcloned_in pPSR177a Name of Probe that a PSR177 piece has been subcloned as Image Survey 2 PSR177 Name of image of autoradiogram or other picture Data_source Sorrells, Mark E. Who provided this information Date 93.12.25 When was the information submitted, in format yy.mm.dd ----------------------------------------------------------------------------- There are a few additional fields that are more complicated, such as Polymorphism and Wheat_polymorphism. The best way to find out about these is to look at some examples. Try the query "find probe wheat_polymorphism".