Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/16: Difference between revisions
| Line 59: | Line 59: | ||
Expected size fragments from EcoRI/PstI digestion: 2114, 189, 53 | Expected size fragments from EcoRI/PstI digestion: 2114, 189, 53 | ||
[[Image:2015-04-16_DBN001_pSB1A3_digest_annotated.png]] | |||
No trace of plasmid, just a smear of (presumably) genomic DNA. Will have to repeat plasmid prep and hope for a higher yield. | |||
<!-- ##### 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. ##### --> | ||
Revision as of 21:03, 16 April 2015
 Project name
|
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||
Plasmid Miniprep of gBlock in mCherry vector--Notes-- Both colonies grown in liquid culture are normal colored (not red). Cell density appears low though, with lots of sticky filaments. Indication that they were not subjected to continuous shaking overnight. Also, for future reference: when performing digestion/ligation, set up a control plate using linearized vector treated with phosphatase. Presence of colonies on this plate indicates that the phosphatase step did not completely work.
--DNA Concentration--
--Restriction Digest--
Digest for 15 minutes at 37°C. NOTE: Should've used SpeI, not PstI. Oh well, won't make much of a difference for analysis of the sequence.
--Gel Electrophoresis--
No trace of plasmid, just a smear of (presumably) genomic DNA. Will have to repeat plasmid prep and hope for a higher yield.
| ||||||||||||||||||||||||
