The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
Dilute primers that came from IDT (10x dilution of concentration)
BL_Primer09 26.6 nm --> 266 μL dilution
BL_Primer10 25.7 nm --> 257 μL dilution
Thaw and dilute primers that came (x10 dilution)
90 μL water
10 μL primer stock
(10 μm in each tube for PCR reaction)
______________________________________
Assemblies
1. hPCD - Pflex - BL01
2. hPCD - BL01
#
Template
Primers
1
hPCD
9, 2
2
Plflex
3, 4
3
BL01
5,10
4
hPCD
9, 7
5
BL01
8,10
μL
Template
.5 μL
Primer 1 (10 μm)
1.0 μL
Primer 2 (10 μm)
1.0 μL
2*GoTaq
12. 5
DH2O
10.0 μL
Run PCR with preset GoTaq settings. Need to ask Dr. Haynes to be certain settings are correct for my reactions.
______________________
*Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
Reagent
Volume
DNA (plasmid)
20.0
10x buffer
3.0
XbaI
1.0
SpeI
1.0
dH2O
5.0
30 μL --> 37°C/ ~10 min.
Vi helped me with running gel because I had class. Vi will put digest into gel and will get results.
I believe that I did not wait long enough after I microwaved the gel to put in the ethnium bromide. I need to be sure that I wait until its cooled down.
Vi cut gel and has it available for me to work on tomorrow afternoon.
Vi also took out my PCR reaction. She told me that the lid was not closed all the way, so I may need to redo rxns.