Journal: PLoS ONE

Article Title: The effects of tissue fixation on sequencing and transcript abundance of nucleic acids from microdissected liver samples of smallmouth bass ( Micropterus dolomieu )

doi: 10.1371/journal.pone.0236104

Figure Lengend Snippet: (A) Mean (+ standard error) of RNA and (B) DNA concentrations (μg/mm 3 ) of microdissected smallmouth bass liver samples fixed in 10% neutral buffered formalin (NBF), Z-Fix ® (ZF), and PAXgene® (PG) for 24 hours, 48 hours, 7 and 14 days. Samples preserved in 95% alcohol (ETOH) and RNALater® were included as controls.

Article Snippet: Tissues preserved in PG were removed from the PAXgene® Tissue FIX (Product # 765312, QIAGEN) after 4 hrs at RT, placed in the PAXgene® Tissue STABILIZER solution (Product # 765512, QIAGEN), and stored at 4°C for 24 hrs, 48 hrs, seven and 14 days.

Techniques:

Journal: PLoS ONE

Article Title: The effects of tissue fixation on sequencing and transcript abundance of nucleic acids from microdissected liver samples of smallmouth bass ( Micropterus dolomieu )

doi: 10.1371/journal.pone.0236104

Figure Lengend Snippet: (A) Mean RIN values of RNA and (B) fragment size (bp) of DNA from samples fixed in 10% neutral buffered formalin (NBF), Z-Fix (ZF) and PAXgene (PG) for 24 hours, 48 hours, 7 and 14 days. Samples preserved in 95% alcohol (ETOH) and RNAlater were included as controls.

Article Snippet: Tissues preserved in PG were removed from the PAXgene® Tissue FIX (Product # 765312, QIAGEN) after 4 hrs at RT, placed in the PAXgene® Tissue STABILIZER solution (Product # 765512, QIAGEN), and stored at 4°C for 24 hrs, 48 hrs, seven and 14 days.

Techniques:

Journal: Acta Naturae

Article Title: A Highly Productive CHO Cell Line Secreting Human Blood Clotting Factor IX

doi:

Figure Lengend Snippet: Analysis of target gene integrity in the genomic DNA and cDNA from producer cell lines by PCR, RT-PCR, Southern blotting and pseudo-Northern blotting. DNA ladder bands are shown as bps. Panel A – amplification of whole ORF regions of the target genes in genomic DNA (gDNA) by PCR. The annealing temperature gradient is shown with red triangles; the linear shift in annealing temperature was from 53 to 68 oC. The expected amplification products of the FIX , furin, and VKORC1 genes are shown with red arrows. Panel B – RT-PCR products for total mRNA with primers towards the 5’- and 3’- ends of the FIX ORF; the amplification product of expected size is shown with a red arrow. Panel C – Southern blotting of gDNA with biotin-labeled probes. The IDbla probe is the probe targeting the IRES and DHFR regions, the ampicillin resistance gene (the bla gene) and the region of plasmid origin of replication (ori). The FIX probe is the probe targeting the FIX ORF region. Sizes of the detected target restriction fragments are shown with arrows. Negative agarose gel images; the membrane images were contrast-enhanced to add visibility. Panel D – Analysis of cDNA by pseudo-Northern blotting. Denotation is the same as that in panel C. The actual position and size of FIX cDNA are shown with an arrow

Article Snippet: The commercially available clone of human FIX cDNA, pCMV6-XL4/NM_000133.2 (sc126517, Origene, USA), and adapter primers AD-9-AbsF and AD-9- NheR ( ) were used as a source of FIX ORF.

Techniques: Reverse Transcription Polymerase Chain Reaction, Southern Blot, Northern Blot, Amplification, Labeling, Plasmid Preparation, Agarose Gel Electrophoresis, Membrane