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==== <u>Library pooling</u> ==== =====<span style="color:mediumblue"><u> 006-T samples</u></span>===== * Since I may need to sequence only one replicate, I combine sequencing libraries into two set; set 1 and set2. Set 1 set the first priority, so I included sample 6-T-2_2 in set one as sample 6-T-2_1had very faint signal. I included sample 6-T_2.1 in set 2 <span style="color:mediumblue">NP-RRBS-6T-set1_100ng-Jun28</span> {| {{table}} border = 1 | align="center" style="width:120px;background:#f0f0f0;"|'''Sample''' | align="center" style="width:60px;background:#f0f0f0;"|'''Index''' | align="center" style="width:80px;background:#f0f0f0;"|'''Conc. (ng/ul)''' | align="center" style="width:80px;background:#f0f0f0;"|'''Volume for 120ng (ul)''' |- | 6-T_1.1||1||6.53||18.36 |- | <span style="color:crimson">6-T_2.2</span>||4||7.14||16.81 |- | 6-T-3.1||5||10.60||11.32 |- | 6-T-4.1||7||6.19||19.39 |- | 6-T-5.1||25||4.31||27.83 |} :- Add 20ul of 6X loading dye and load 37.9ul in 3 well of 5-well 6% TBE gel <span style="color:mediumblue">NP-RRBS-6T-set2_100ng-Jun28</span> * Set 2 I include 4 libraries and do PAGE size selection of sample 6-T-4.2 separately since it had very faint smear. {| {{table}} border = 1 | align="center" style="width:120px;background:#f0f0f0;"|'''Sample''' | align="center" style="width:60px;background:#f0f0f0;"|'''Index''' | align="center" style="width:80px;background:#f0f0f0;"|'''Conc. (ng/ul)''' | align="center" style="width:80px;background:#f0f0f0;"|'''Volume for 100ng (ul)''' |- | 6-T-1.2||2||9.91||10.09 |- | 6-T-2.1||23||2.00||49.90 |- | 6-T-3.2||6||6.70||14.93 |- | 6-T-5.2||10||8.26||12.11 |} :- Add 20ul of 6X loading dye and load 35.68 in 3 well of 5-well 6% TBE gel <span style="color:mediumblue">NP-RRBS-6T-4_2-set2_100ng-Jun28</span> {| {{table}} border = 1 | align="center" style="width:120px;background:#f0f0f0;"|'''Sample''' | align="center" style="width:60px;background:#f0f0f0;"|'''Index''' | align="center" style="width:80px;background:#f0f0f0;"|'''Conc. (ng/ul)''' | align="center" style="width:80px;background:#f0f0f0;"|'''Volume (ul)''' |- | 6-T-4.2||8||0.71||50.00 |} :- Add 10ul of 6X loading dye and load 60ul in 1 well of 5-well 6% TBE gel =====<span style="color:red"><u> 007-T samples</u></span>===== <span style="color:red">NP-RRBS-7T-set1_100ng-Jun28</span> {| {{table}} border = 1 | align="center" style="width:120px;background:#f0f0f0;"|'''Sample''' | align="center" style="width:60px;background:#f0f0f0;"|'''Index''' | align="center" style="width:80px;background:#f0f0f0;"|'''Conc. (ng/ul)''' | align="center" style="width:80px;background:#f0f0f0;"|'''Volume for 100ng (ul)''' |- | 7-T-1.1||13||5.12||19.55 |- | 7-T-2.1||15||4.18||23.93 |- | 7-T-3.1||27||2.70||36.98 |- | 7-T-4.1||19||9.30||10.75 |- | 7-T-5.1||21||5.76||17.35 |} :- Add 25ul of 6X loading dye and load 44.52ul in 3 well of 5-well 6% TBE gel <span style="color:red">NP-RRBS-7T-set2_100ng-Jun28</span> {| {{table}} border = 1 | align="center" style="width:120px;background:#f0f0f0;"|'''Sample''' | align="center" style="width:60px;background:#f0f0f0;"|'''Index''' | align="center" style="width:80px;background:#f0f0f0;"|'''Conc. (ng/ul)''' | align="center" style="width:80px;background:#f0f0f0;"|'''Volume for 100ng (ul)''' |- | 7-T-1.2||14||3.65||27.41 |- | 7-T-2.2||16||6.55||15.27 |- | 7-T-3.2||18||3.72||26.89 |- | 7-T-4.2||20||3.59||27.82 |- | 7-T-5.2||22||3.82||26.20 |} :- Add 30ul of 6X loading dye and load 51.2ul in 3 well of 5-well 6% TBE gel<br>
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