Matt:LabNotes/2013-7-11: Difference between revisions

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>Mzcai
(Created page with "Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-7-9 ====Amplification with Sequencing Adapters==== *Common linker sequence in every probe i...")
 
>Mzcai
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*Common linker sequence in every probe is: CTTCAGCTTCCCGATATCCGACGGTAGTGT (Porecca et al. 2007)
*Common linker sequence in every probe is: CTTCAGCTTCCCGATATCCGACGGTAGTGT (Porecca et al. 2007)
**[[Media:probe2padlockFISSEQ_2012_0bp.txt|./probe2padlockFISSEQ_2012_0bp.pl]]  
**[http://genome-tech.ucsd.edu/LabNotes/index.php/Capture_Test_of_CES36k18bp Capture Test of CES36k18bp]
**[[Media:probe2padlockFISSEQ_2012_20bp.txt|./probe2padlockFISSEQ_2012_20bp.pl]]
**[[Media:probe2padlockFISSEQ_2012_20bp.txt|./probe2padlockFISSEQ_2012_20bp.pl]]


*[http://genome-tech.ucsd.edu/LabNotes/index.php/Illumina_GA/Oligo_info#Primers_for_LC_Sciences_library-free_protocol primers]
*[http://genome-tech.ucsd.edu/LabNotes/index.php/Illumina_GA/Oligo_info#Primers_for_LC_Sciences_library-free_protocol Primers for Agi26k]
*Primers for CES36k18bp: AmpF6.3NH2 and AmpR6.3NH2


{| {{table}} border = 1
{| {{table}} border = 1
Line 16: Line 17:
| 1||NTC-Agi26k_20gap||Indx7
| 1||NTC-Agi26k_20gap||Indx7
|-
|-
| 2||gDNA-Agi26k_20gap||Indx45
| 2||gDNA-Agi26k_20gap||Indx77
|-
|-
| 3||cDNA-Agi26k_20gap||Indx76
| 3||cDNA-Agi26k_20gap||Indx78
|-
|-
| 4||gDNA-CES36k18bp||Indx77
| 4||gDNA-CES36k18bp||N/A
|} <br>
|} <br>



Revision as of 21:14, 11 July 2013

Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-7-9

Amplification with Sequencing Adapters

Tube # Sample Index#
1 NTC-Agi26k_20gap Indx7
2 gDNA-Agi26k_20gap Indx77
3 cDNA-Agi26k_20gap Indx78
4 gDNA-CES36k18bp N/A


Components 1X Reaction 5X Reaction
Captured Template 1 0
10uM Forward + Indx 0.4 0
10uM Reverse 0.4 2.0
2X KAPA SYBG FAST MM 12.5 62.5
H2O 10.7 53.5
Total 25 125
 Program
 *98°C 30sec -> (98°C 10sec -> 52°C 30sec -> 72°C 30sec) x8 cycles -> (98°C 10sec -> 72°C 30sec) x16 cycles -> 72°C 3 min -> 15°C hold