User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/06/24

From OpenWetWare

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

Contents

Re-Amp Alien Gene LongPCR

Re-amp Mix 19 and 21

' [CONC] 1X 2 ' ' AMP Lane Template MIX Contents ' Size Fosmid
Water10.621.219CCA3111AV10106 FWDAV10108.2 RVS13400AV_TEL_L_B
Takara LA Taq5U0.20.421CCA355AV10108 FWDAV10111 RVS14846AV_TEL_L_B
Takara LA Buffer10X24
dNTP2.5uM each3.26.4
*Template1ng24
Primer Mix5uM each2
10.0ul3618.0ul/RXN
add 2.0 Primer Mix
* CCA3
PCR Cycle
94C 1 min
98C10 sec
65C15 min
30X

load 2.0 ul of sample

Image:24June10.jpg

Library preparation for Alien genes

Confirmation of adapter ligation by PCR

Reaction mixture conc. 1x x4 ' lane Template Primer set
Template (Adapter ligated AGM fragments, purified by MinElute)0.25112 days ligation (21.9 ng/ul)Primer Atitin + Btitin
TopTaq Buffer10x1422 days ligation (21.9 ng/ul)Primer Atitin
dNTP mix10 mM each0.20.833 days ligation (31.0 ng/ul)Primer Atitin + Btitin
Primer mix5 uM each0.543 days ligation (31.0 ng/ul)Primer Atitin
TopTaq5 U/ul0.050.2
Milli Q832
10 ul
PCR cycle
94C3 min
98C10 sec
60C3 min
x30
72C5 min

Image:0131-2.jpg

Personal tools