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=== Below is a rendering of the page up to the first error. === </parsererror> [[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)== 17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)[[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)=== [[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)== Date:2/7/2018 '''Figure Description''' PCR of different Taq enzymes {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Template | 2X PCR Mix | Primer |- | 1 | 3011 H(+) | DreamTaq | pMini T2 |- | 2 | 3011 H | DreamTaq | pMIniT2 |- | 3 | 3011 H | PCR MIx | pMIniT2 |- | 4 | 3011 H | 0.1Taq | pMIniT2 |- | 5 | 3011 H | DreamTaq | 3129 F&R |} '''Results: We didn't get good results.''' [[File:Test of different products.JPG|RTENOTITLE]] '''Figure Despriction''' {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Template | 2X PCR | Primer |- | 1 | 3011 H | DreamTaq | 3129 F&R |- | 2 | 3011 H | DreamTaq | pMiniT2 |- | 3 | 3011 H | PCR Mix | pMiniT2 |- | 4 | 3011 L | 0.1 Taq | pMiniT2 |- | 5 | 3011 L | PCR Mix | 3130 F&R |- | 6 | 3011 L | 0.1 Taq | 3130 F&R |} [[File:PCR of different enzymes 2.JPG|RTENOTITLE]] '''Figure Despriction: ''' '''Annealing:55oC, Extension:30s,''' {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Template | 2X PCR Mix | Primers |- | 1 | 3011 L | phusion | pMini T2 F&R |- | 2 | <div>3011 L</div> | phusion | pMini T2 F&R<br/> |- | 3 | 3011 L<br/> | phusion | pMini T2 F&R<br/> |- | 4 | 3011 L<br/> | phusion | pMini T2 F&R<br/> |- | 5 | 3011 L | kfu | pMini T2 F&R<br/> |- | 6 | 3011 L | kfu | pMini T2 F&R<br/> |- | 7 | 3011 L<br/> | kfu | pMini T2 F&R<br/> |- | 8 | 3011 L<br/> | kfu | pMini T2 F&R<br/> |- | 9 | 3011 L<br/> | KOD | pMini T2 F&R<br/> |- | 10 | 3011 L<br/> | KOD | pMini T2 F&R<br/> |- | 11 | 3011 L<br/> | KOD | pMini T2 F&R<br/> |- | 12 | 3011 L | KOD | pMini T2 F&R<br/> |- | 13 | 3011 L | 0.5ul Taq+ 0.5ul pfu | pMini T2 F&R<br/> |- | 14 | 3011 L | 0.5ul Taq+ 0.5ul pfu<br/> | pMini T2 F&R<br/> |- | 15 | 3011 L(-) | no DNA polymerase | pMini T2 F&R<br/> |} [[File:Polymerase QC for using PCR.JPG|RTENOTITLE]] '''Results: The enzymes may not work.''' [[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)== 17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)[[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST) [[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)== '''Date:2/8/2018''' '''Figure Despriction''' '''Digestion of 3011 L_3011 H''' http://www.excellgen.net/eln/images/3/3e/Digestion_of_3011_L_3011_H.JPG '''Result:#1,#2,#4,#5,#7,#8 has bands. So we did PCR for these samples.''' '''Figure Despriction''' {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Tempate | 2X PCR Mix | Primer |- | 1 | 3011 L (For 3130) | 0.1 Taq | pMiniT2 F&R |- | 2 | 3011 L | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 4 | 3011 L | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 5 | 3011 L | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 6 | 3011 H(For 3129) | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 7 | 3011 H | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 8 | 3011 H | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 9 | 3011 H | 0.1 Taq<br/> | pMiniT2 F&R<br/> |- | 10 | 3011 H(+) | 0.1 Ta1 |} [[File:PCR of Mini Prep for 3130 and 3129.JPG|RTENOTITLE]] Result: we picked #1,#8 to do sequencing. #1 is no primering.#8 is missing C in Kpn I. Friday,2/9/2018, we ordered another sequencing for #4. '''Figure Despriction''' '''Subclone of 3009''' Subclone from 3009-4-pMiniT2, Vector from 2492 [[File:Subclone 3009-4 2492.JPG|RTENOTITLE]] Result:cut the bands and do ligation. [[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)== 17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)17:16, 13 February 2018 (EST)=== [[User:Paul.xu|Paul.xu]] ([[User talk:Paul.xu|talk]]) 17:16, 13 February 2018 (EST)== Date:2/9/2018 '''Protein purfication: 15 samples.''' '''Use buffer B to elute''' '''Concentration:''' {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Concentration |- | 1 | -0.53 |- | 2 | 0.48 |- | 3 | 1.59 |- | 4 | 2.24 |- | 5 | 1.26 |- | 6 | 1.87 |- | 7 | 1.17 |- | 8 | 0.63 |- | 9 | 0.50 |- | 10 | 0.30 |- | 11 | 0.53 |- | 12 | 0.29 |- | 13 | 0.14 |- | 14 | 0.11 |- | 15 | 0.09 |} '''PCR Gene synthesis -----3010''' {| border="1" cellspacing="1" cellpadding="1" style="width: 778px;" |- | # | Tempate | style="width: 111px;" | Oligo | style="width: 174px;" | 2X PCR Mix | style="width: 46px;" | Primer | style="width: 307px;" | Note |- | 1 | 3135-4 | style="width: 111px;" | 3010-D1,D2,D3<br/> | style="width: 174px;" | Platinum 2X SuperMix HiFi | style="width: 46px;" | C4,C5,C6 | style="width: 307px;" | C4--10uM,C5,C6,D1,D3--0.5uM,D3--10uM |- | 2 | 3135-4 <br/> | style="width: 111px;" | 3010-D1,D2,D3<br/> | style="width: 174px;" | Platinum 2X SuperMix HiFi<br/> | style="width: 46px;" | C4,C5,C6<br/> | style="width: 307px;" | C4--10uM,C5,C6,D1,D3--0.5uM,D3--10uM |- | 3 | 3135-4 <br/> | style="width: 111px;" | 3010-D1,D2,D3<br/> | style="width: 174px;" | Platinum 2X SuperMix HiFi<br/> | style="width: 46px;" | C4,C5,C6<br/> | style="width: 307px;" | C4--10uM,C5,C6,D1,D3--0.5uM,D3--10uM<br/> |- | 4 | 3135-4 (+) | style="width: 111px;" | 3010-C6,D1<br/> | style="width: 174px;" | Platinum 2X SuperMix HiFi<br/> | style="width: 46px;" | C6.D1 | style="width: 307px;" | C6,D1---10uM |} [[File:PCR of 3010 gene synthesis.JPG|RTENOTITLE]] '''PCR Gene synthesis -----3001''' {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Oligo | 2X PCR Mix | Primer | Note |- | 1 | 3001 Oligo Mix | Q5 | E6,F5 | <br/> |- | 2 | 3001 Oligo Mix<br/> | Q5 | E6,F5 | <br/> |- | 3 | 3001 Oligo Mix<br/> | Q5 | E6,F5 | <br/> |- | 4 | 3001 Oligo Mix<br/> | Q5 | E6,F5 | <br/> |} <div></div [[File:PCR of 3001 gene synthesis.JPG|RTENOTITLE]] '''Result: we didn't synthesiz ''''''3001'''''' successfully.''' '''Colony screening for 3009 subclone''' '''Figure Desperction''' {| border="1" cellspacing="1" cellpadding="1" style="width:500px;" |- | # | Template | 2X PCR Mix | Primer |- | 1 | 3009-1 | 0.1Taq | pMiniT2 F&R |- | 2 | 3009-2 | ..... | ............ |- | 3 | 3009-3 | ..... | ............. |- | 4 | 3009-4 | ..... | .............. |- | 5 | 3009-5 | ..... | .............. |- | 6 | 3009-6 | ..... | ............. |- | 7 | 3009-7 | ..... | ............. |- | 8 | 3009-8 | ..... | .............. |- | 9 | 3009-9 | ..... | ................. |- | 10 | 3009-10 | ..... | .................... |- | + | 3011 L | 0.1Taq | pMiniT2 F&R |} [[File:Colony screening.JPG|RTENOTITLE]] </div>[[Category:My ELN]]
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