1.1.2 主要酶和试剂 Taq酶、限制性内切酶EcoR I、Not I、T4 DNA连接酶、DNA marker和Prime STAR② Max DNA Polymerase均购自TaKaRa公司。脂质体Lipofectamine 2000购自Invitrogen公司。小提质粒试剂盒和胶回收试剂盒购自宝生物工程大连有限公司。 1.2 方法 1.2.1 SREBP-1c基因的提取及鉴定 按照SREBP-1c基因序列(...
Then, the cDNA/DNA hybrid was purified with AMPureXP and amplified with PrimerSTAR GXL DNA polymerase (Takara Bio) using PCR primers (fwd_CTACACTCGTCGGCAGCGTC, rev _GAGATGTCTCGTGGGCTCGG) for 7 cycles. The library was then subjected to the SQK-LSK110 (Oxford Nanopore Technologies) ...
1.1.2 主要试剂及培养基 PrimeSTAR Max Premix DNA 聚合酶购于日本 TaKaRa 公司 ;DreamTaq DNA 聚合酶,碱性磷酸酶及限制性内切酶购于赛默飞公 司 ;T4 DNA 连接酶购于 NEB 公司 ;Phanta Max Mix DNA 聚合酶购于南京诺唯赞生物科技股份有限公 司.乳糖及岩藻糖购于 Sigma 公司 ;GDP- 岩藻糖及 荧光乳糖由...
To generate pCDF2::3HA-CDF2, the CDF2 promoter and the full-length coding region were amplified from Col-0 genomic DNA and 3HA was amplified from plasmid pALLIGATOR2 using PrimeSTAR GXL DNA Polymerase (Takara Bio), then overlap PCR was performed using CDF2 promoter, 3HA and CDF2 coding...
PrimeSTAR GXL DNA polymeraseTakaraCat# R050A ProtoScript II Reverse TranscriptaseNew England BiolabsCat# M0368S NEBNext® Ultra™ II Non-Directional RNA Second Strand Synthesis ModuleNew England BiolabsCat# E6111S/L SureSelectXT HS and XT Low input enzymatic fragmentation kitAgilentCat# 5191-4080...
To generate recombinant SARS-CoV-2, the nine pmW118 plasmid vectors were subjected to amplification of the cDNA fragments (F1–F9-10) of SARS-CoV-2 XBB.1 and XBB.1.5 by PrimeSTAR GXL DNA polymerase (Takara) with the primer sets26. To generate mutant plasmids bearing mutations (S:S486P ...
PrimeSTAR® Max DNA Polymerase TaKaRa Cat# R045A SuperScript III First-Strand Synthesis SuperMix Invitrogen Cat# 18080400 RNase H Thermo Scientific Cat# EN0201 Lipofectamine® 3000 Transfection Kit Invitrogen Cat# L3000075 CellMask™ Deep Red Plasma membrane Stain Invitrogen Cat# C10046 DAPI Th...
Furthermore, these results suggest that other K36MTs may prime gene bodies with H3K36me1 in advance of SETD2 activity, which is tightly associated with transcription. Intergenic H3K36me2 is predominantly deposited by NSD1 and in part by NSD2 in mMSCs...
Cells (50 × 103) were washed in PBS and spotted on Superfrost Plus microscope slides (Thermo Fisher Scientific, #10149870) using a cytospin centrifuge (CR2000 Small Prime Centrifuge, Centurion). Fixation was carried out for 10 min in PBS, 4% paraformaldehyde (Thermo Fisher Scientific, #2...
(T)12VN primer was used to prime the reverse transcription reaction before PCR amplification with a gene-specific forward primer (indicated in B) and an additional priming sequence contained within the RT primer for 30 cycles. Usage of the AAUAAAA sites marked in B is expected to lead to ...