The PCR product size ranged from 110 to 274 bp. All the primers produced amplicons in agreement with the expected sizes. Most of the SSR primers (139 primer pairs) showed significant deviation from HW equilibrium (P < 0.05). Partial correlation analysis showed that significant positive ...
However, the time window for positive RT-PCR viremia is relatively short, roughly three to seven days, thus a suspected ZIKV should not be regarded as a negative case, which requires IgM tests for further confirmation [69]. Therefore, to avoid excluding the part of positive ZIKV cases in ...
Reverse transcription-PCR analysis (RT-PCR) was conducted to assess TRV infection and its systematic spread. We extracted total RNA from a 50–100 mg leaf tissue sample using the ExtractRNA kit (Evrogen, Moscow, Russia), based on TRIzol reagent. RT-PCR and qPCR analyses were performed with ...
A ligation reaction between pCR2.1 and the target gene was set up with a 1:1 vector/insert molar ratio. The ligation reaction was carried out in 5X T4 DNA ligase reaction buffer at 25 °C for 1 h. Two microliters of the ligation reaction was used to transform 50 μl of One Shot...
The AND gate Reaction A utilizes New England Biolabs OneTaq Polymerase (Catalog No. M0480) PCR recommendations. However, instead of running the reaction for 30 cycles, the AND gate only requires one cycle. Each AND gate reaction has a final volume of 25μL and includes 5μL of OneTaq 5X...
unless this site was present in the insert sequence. Positive clones were screened by colony PCR using one primer that binds to the vector and a second primer that binds to the insert. All constructs were verified by sequencing. Detailed information on all plasmids used in this study is provid...
Each PCR product was purified, A-tailed, and ligated into a predigested pGEM-T vector as described previously (23). After sequence verification of each cassette, (protein L)5 was constructed by sequential replacement of each I27 cassette in (C47S C63S I27)5(24) with its analogous protein...
3A). This was achieved by using positive PCR products from the same 2–6 or 1–5 primer pairs described previously and sequencing these PCR products with either primer 1 or 6 depending on the initial primer pairs used (Fig. 3A). In this case, the sequencing results informed us as to ...
The SSR primer pairs were designed using the Oligo 7.0 program46based on the following parameters: 1) primer length of 17–25 bp with 20 bp as the optimum; 2) PCR product size ranging from 100 to 500 bp; 3) GC content of 40–70% with an optimum of 50%; 4) melting temperat...
To further support immunostaining data, quantitative PCR was performed on sub-confluent and differentiated cells from P2 and P6. Briefly, RNA was harvested with the RNEasy Mini Kit (Qiagen #74104, Hilden, Germany) and cDNA was prepared using 200 ng of RNA for each reaction with the iScript...