User:Alexander S Mikheyev/Notebook/rotifer alien genes/2009/12/11

From OpenWetWare

Jump to: navigation, search
Alien genes Main project page
Previous entry      Next entry

Contents

Cycle Sequencing Clones M13FWD

83 colonies PCR amped -> sequencing

Nanodrop Data

Sample ID Nucleic Acid Conc. Unit 260/280 260/230
clone120.6ng/µl1.591.1
clone212.8ng/µl1.931.12
clone38.8ng/µl1.461.21
clone410.2ng/µl1.571.1
clone521.9ng/µl1.491.14
Cycle Sequencing ' ' '
ReagentsConcentration1X87X
BDT0.543.5
M13 FWD0.434.8
DMSO0.18.7
5X Seq Buffer5X0.434.8
H203.6313.2
PEG DNA variable1
TOTAL6435
Cocktail per well = 5.0ul
PCR 96C10 sec
50C5 sec
60C4 min
60 cycles

Cycle Sequencing AV10007, AV10016, AV10027, AV10153, PR10008 and 5475F + 8047R

Cycle Sequencing ' ' '
ReagentsConcentration1X14X
BDT0.57
DMSO0.11.4
5X Seq Buffer5X0.45.6
H203.650.4
*Primer15uM0.4
*PEG DNA variable1
TOTAL664.4
Cocktail per well = 4.6ul
*add primer and PEG separately
PCR 96C10 sec
50C5 sec
60C4 min
60 cycles
PrimersSequence FWD and RVS
AV10007
AV10016
AV10027
AV10153
PR10008
5475F + 8047R
Personal tools