Hiscript q select rt supermix for pcr

Manufactured by Vazyme

Sourced in China

HiScript Q Select RT SuperMix for PCR is a reverse transcription (RT) reagent kit designed for cDNA synthesis from RNA templates. It includes an optimized buffer, reverse transcriptase enzyme, and RNase inhibitor to facilitate efficient and reliable cDNA production for subsequent PCR amplification.

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Lab products found in correlation

Total rna kit, by Tiangen Biotech (2 mentions)

Close spelling variants of this product

HiScript II Q Select RT SuperMix for qRT-PCR HiScript Q Select RT SuperMix HiScript Q RT SuperMix

2 protocols using hiscript q select rt supermix for pcr

Chicken Spleen-Derived scFv Generation

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The spleen tissue from the hen with the highest IgY titer was collected to extract the total RNA by using Total RNA Kit (Tiangen Biotech, Beijing, China), and the first-strand cDNA was synthesized by HiScript Q Select RT SuperMix for PCR (Vazyme Biotech, Nanjing, China). The heavy variable fragment (V_H) and light chain variable fragment (V_L) genes were amplified by PCR with primers HF-Sfi I and HR-Linker, LF-Linker and LR-Not I, respectively (Table 1). The V_H and V_L which both contained the sequence of a peptide linker were assembled to scFv with primers HF-Sfi I and LR-Not I by Overlap PCR.Table 1

Primers used for PCR

Name	Primer sequences (5ʹ–3ʹ)	Application
HF-Sfi I	ATGTCTATGGCCCAGCCGGCCGTGACGTTGGACG	V_H
HR-Linker	CAGAGCCACCTCCGCCTGAACCGCCTCCACCGGAGGAGACGATGACTTCGG	V_H
LF-Linker	TTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGCGCTGACTCAGCCGTCCT	V_L
LR-Not I	AGTTACTGGAGCGGCCGCACCTAGGACGGTCAGGG	V_L

Ge S., Wu R., Zhou T., Liu X., Zhu J, & Zhang X. (2022). Specific anti-SARS-CoV-2 S1 IgY-scFv is a promising tool for recognition of the virus. AMB Express, 12, 18.

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Construction of Chicken ScFv Antibody

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The hen’s spleen was collected to extract the total RNA by Total RNA Kit (Tiangen Biotech, Beijing, China), and the first-strand cDNA was synthesized by HiScript Q Select RT SuperMix for PCR (+gDNA wiper, Vazyme Biotech, Nanjing, China). The heavy variable fragment (V_H) and light chain variable fragment (V_L) genes were amplified by PCR with primers (Table 1). The V_H and V_L were assembled with primers HF-EcoR I & LR-Hind III by Overlap PCR. The products of overlap PCR (scFv) were purified through Gel Extraction Kit (Omega, Norcross, GA, USA). This protocol was performed as described in [14 (link)].Table 1

Primers used for PCR.

Name	Primer sequences (5′-3′)	Application
HF-EcoR I	CGGAATTCGGCCGTGACGTTGGACG	V_H
HR-Linker	CAGAGCCACCTCCGCCTGAACCGCCTCCA	V_H
HR-Linker	CCGGAGGAGACGATGAC	V_H
LF-Linker	TTCAGGCGGAGGTGGCTCTGGCGGTGGCG	V_L
LF-Linker	GATCGGCGCTGACTCAGCCGTCCT	V_L
LR-Hind III	AATAAGCTTACCTAGGACGGTCAGGG	V_L

Ge S., Xu L., Li B., Zhong F., Liu X, & Zhang X. (2020). Canine Parvovirus is diagnosed and neutralized by chicken IgY-scFv generated against the virus capsid protein. Veterinary Research, 51, 110.

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