User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/04: Difference between revisions
(Autocreate 2013/01/04 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing) |
(fix raw html notebook nav) |
||
| (4 intermediate revisions by one other user not shown) | |||
| Line 2: | Line 2: | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==Reconciling information and thoughts after break== | ||
I did some reading and thinking and research during the xmas break. This is the result. | |||
=Housekeeping Genes= | |||
Finally found some housekeeping primers: | |||
18S-F908 18S a AGCGAAAGCATTTGCCAAGA 401 This study (SVB) | |||
18S-R1309 18S a AGTCTCGTTCGTTATCGGAATT This study (SVB) | |||
(from http://wwwnc.cdc.gov/eid/article/11/5/05-0211-t1.htm) | |||
=Temps and new primers= | |||
P2/P8 primers work best at 60C, confirming my lab work. | |||
(from http://www.academia.edu/1460726/Sex_Identification_of_Some_Pet_Birds_by_Polymerase_Chain_Reaction-Based_Methods ) | |||
also, this paper suggests using some new primers and was getting results very similar to my ones. Going to try and order NP an MP primers. See this paper for what a positive and negative result looks like using P2/P8 primers. | |||
NP primer: | |||
5’-GAG AAA CTG TGC AAAACA G-3’ | |||
MP primer: | |||
5’-AGT CAC TAT CAG ATC CGG AA-3’ | |||
Also this page suggests using 1μL of 10μM primer to give final concentration of 0.5μM in 20μL. Maybe I should start running the reactions at 20ul instead. | |||
Mushy primers: | |||
ITS1 5'-TCCGTAGGTGAACCTGCGG-3' forward all | |||
ITS4 5'-TCCTCCGCTTATTGATATGC-3' reverse all | |||
'''EDIT THESE PAGES TO SEE NICE FORMATTING''' | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | ||
|} | |} | ||
__NOTOC__ | __NOTOC__ | ||
Latest revision as of 22:20, 26 September 2017
 Project name
|
 Main project page Previous entry Next entry
|
Reconciling information and thoughts after breakI did some reading and thinking and research during the xmas break. This is the result. Housekeeping GenesFinally found some housekeeping primers: 18S-F908 18S a AGCGAAAGCATTTGCCAAGA 401 This study (SVB) 18S-R1309 18S a AGTCTCGTTCGTTATCGGAATT This study (SVB) (from http://wwwnc.cdc.gov/eid/article/11/5/05-0211-t1.htm) Temps and new primersP2/P8 primers work best at 60C, confirming my lab work. (from http://www.academia.edu/1460726/Sex_Identification_of_Some_Pet_Birds_by_Polymerase_Chain_Reaction-Based_Methods ) also, this paper suggests using some new primers and was getting results very similar to my ones. Going to try and order NP an MP primers. See this paper for what a positive and negative result looks like using P2/P8 primers. NP primer: 5’-GAG AAA CTG TGC AAAACA G-3’ MP primer: 5’-AGT CAC TAT CAG ATC CGG AA-3’ Also this page suggests using 1μL of 10μM primer to give final concentration of 0.5μM in 20μL. Maybe I should start running the reactions at 20ul instead.
EDIT THESE PAGES TO SEE NICE FORMATTING | |
