Mus musculus
Protocols: BALB/c mice were immunized with inactivated influenza A virus plus liposomal formulation of 1V270 (TLR7 agonist) and 2B182C (TLR4 agonist) on days 0 and 21. Mice were sacrificed on day 28 and lymphocytes in the draining lymph nodes in the injected sides (inguinal lymph nodes) were collected for BCR sequencing. Total RNA was extracted from lymphocytes using RNeasy Mini Kit (Qiagen, Hilden, Germany) and the quality of RNA was confirmed by Agilent 2200 TapeStation System (Agilent, Santa Clara, CA). Total RNA was converted to complementary DNA (cDNA) with Superscript III reverse transcriptase (Invitrogen, Carlsbad, CA). Then, double strand (ds)-cDNA was synthesized and an P20EA/P10EA adaptor was ligated to the 5' end of the ds-cDNA and then cut with SphI restriction enzyme. PCR was performed with KAPA HiFi DNA Polymerase (Kapa Biosystems, Woburn, MA) using P20EA and IgG constant region-specific primer mCG1 (GACAGGGMTCCAKAGTTCC). The second PCR was performed with P20EA and mCG2 (ACYGRCTCAGGGAARTAVCC) using the same PCR conditions. After Tag PCR amplification with mCG-ST1-R (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCYTTGACMAGGCAYCC) and P22EA-ST1-R, index (barcode) sequences were added by amplification with Nextera XT index kit v2 setA (illumina, San Diego, CA).
ENA-FIRST-PUBLIC
2021-03-01T12:16:03Z
ENA-LAST-UPDATE
2020-03-09T15:45:59Z
External Id
SAMEA6615251
INSDC center name
Moores Cancer Center, University of California San Diego, La Jolla 92093, United States
INSDC first public
2021-03-01T12:16:03Z
INSDC last update
2020-03-09T15:45:59Z
INSDC status
public
Submitter Id
E-MTAB-8870:Sample 13_USH139G
antigen/inactivated influenza a virus
10 ug/injection
broker name
ArrayExpress
common name
house mouse
day harvested samples
day 28
days vaccinated
days 0 and 21
genotype
wild type genotype
organism part
inguinal lymph node
sample name
E-MTAB-8870:Sample 13_USH139G
scientific_name
Mus musculus
sex
female
strain
BALB/c