Calendar/2012-7-13: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 (Created page with "==Amplification of Dr. Yi Zhang's DiP libraries Sample== ==qPCR with N2 Barcodes== {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXC...") |
>Alan6017518 |
||
Line 2: | Line 2: | ||
==qPCR with N2 Barcodes== | ==qPCR with N2 Barcodes== | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> | {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> | ||
|- style="font-size: | |- style="background-color:#FCD5B4;font-size:8pt;font-weight:bold" align="center" | ||
|style="text-decoration:underline" width="76" height="15" | DIP Sample | |style="text-decoration:underline" width="76" height="15" | DIP Sample | ||
| width="65" | N2 Barcode | | width="65" | N2 Barcode | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
|style="text-decoration:none" height="15" | shCONT-input | |style="text-decoration:none" height="15" | shCONT-input | ||
| align="center" | 1 | | align="center" | 1 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shCONT-IgG | | height="15" | shCONT-IgG | ||
| align="center" | 2 | | align="center" | 2 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shCONT-5mC | | height="15" | shCONT-5mC | ||
| align="center" | 3 | | align="center" | 3 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shCONT-5hmC | | height="15" | shCONT-5hmC | ||
| align="center" | 4 | | align="center" | 4 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shCONT-5fc | | height="15" | shCONT-5fc | ||
| align="center" | 5 | | align="center" | 5 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shCONT-5caC | | height="15" | shCONT-5caC | ||
| align="center" | 6 | | align="center" | 6 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shTDG-input | | height="15" | shTDG-input | ||
| align="center" | 7 | | align="center" | 7 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shTDG-IgG | | height="15" | shTDG-IgG | ||
| align="center" | 8 | | align="center" | 8 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shTDG-5mC | | height="15" | shTDG-5mC | ||
| align="center" | 9 | | align="center" | 9 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shTDG-5hmC | | height="15" | shTDG-5hmC | ||
| align="center" | 10 | | align="center" | 10 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shTDG-5fC | | height="15" | shTDG-5fC | ||
| align="center" | 11 | | align="center" | 11 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | shTDG-caC | | height="15" | shTDG-caC | ||
| align="center" | 12 | | align="center" | 12 | ||
Line 56: | Line 56: | ||
|} | |} | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> | {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> | ||
|- style="background-color:#B7DEE8;font-size: | |- style="background-color:#B7DEE8;font-size:8pt;font-weight:bold" align="center" | ||
|style="text-decoration:underline" width="115" height="15" | Content | |style="text-decoration:underline" width="115" height="15" | Content | ||
| width="110" | Volume Per Reaction | | width="110" | Volume Per Reaction | ||
| width="98" | MM | | width="98" | MM | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
|style="text-decoration:none" height="15" | Template | |style="text-decoration:none" height="15" | Template | ||
| align="center" | 10 | | align="center" | 10 | ||
| Do not add to MM | | Do not add to MM | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | 5uM PCR F+R Barcode | | height="15" | 5uM PCR F+R Barcode | ||
| align="center" | 1 | | align="center" | 1 | ||
| Do not add to MM | | Do not add to MM | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | 50X SYBG | | height="15" | 50X SYBG | ||
| align="center" | 0.2 | | align="center" | 0.2 | ||
| align="center" | 2.64 | | align="center" | 2.64 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | 2X PhusionHF MM | | height="15" | 2X PhusionHF MM | ||
| align="center" | 25 | | align="center" | 25 | ||
| align="center" | 330 | | align="center" | 330 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | H20 | | height="15" | H20 | ||
| align="center" | 13.8 | | align="center" | 13.8 | ||
| align="center" | 182.16 | | align="center" | 182.16 | ||
|- style="font-size: | |- style="font-size:8pt" align="center" | ||
| height="15" | Total Volume | | height="15" | Total Volume | ||
| align="center" | 50 | | align="center" | 50 |
Revision as of 17:16, 13 July 2012
Amplification of Dr. Yi Zhang's DiP libraries Sample
qPCR with N2 Barcodes
DIP Sample | N2 Barcode |
shCONT-input | 1 |
shCONT-IgG | 2 |
shCONT-5mC | 3 |
shCONT-5hmC | 4 |
shCONT-5fc | 5 |
shCONT-5caC | 6 |
shTDG-input | 7 |
shTDG-IgG | 8 |
shTDG-5mC | 9 |
shTDG-5hmC | 10 |
shTDG-5fC | 11 |
shTDG-caC | 12 |
Content | Volume Per Reaction | MM |
Template | 10 | Do not add to MM |
5uM PCR F+R Barcode | 1 | Do not add to MM |
50X SYBG | 0.2 | 2.64 |
2X PhusionHF MM | 25 | 330 |
H20 | 13.8 | 182.16 |
Total Volume | 50 | 660 |
- Add 20ul 10uM N2-bar-code primers into strip tube
- Add 20ul 10uM PCR_F primers and mix
- We have 5uM F+R N2 Ind primer (Store at 4C Alice yellow 0.2ul tube rack)
- Add 10ul template (for low volume sample add 5ul sample and add 5ul h2o) and 1ul barcode primers (using multi channel pipette) to strip tube
- Add 39ul MM to each well
98C 30sec (98C 10sec -> 63C 20sec-> 72C 30sec) x 20 cycles 72C 2min 15C hold.