Sez6l (clone ID 30362651), Syt1 (clone ID 5363062) cDNAs were obtained GE Dharmacon. Gad1 cDNA (nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (# 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using following primers: F: TGTGCCCAAACTGGTCCT; R: TGGCCGATGATTCTGGTT (Integrated DNA Technologies), gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. Riboprobes against Gad1, Syt1 and Sez6l mRNAs were generated as described previously (Monavarfeshani et al. 2017a (link)). ISH was performed on 16 μm PFA-perfused coronally cryosectioned brain tissue (n = 3 mice) prepared as described above. Tissues were prepared and hybridized at 60°C as previously described (Monavarfeshani et al. 2018 (link), Su et al. 2010 (link)). Bound riboprobes were detected by either horseradish peroxidase (POD)-conjugated anti-DIG or anti-fluorescent antibodies (Roche #: 11426346910 and 11207733910), followed by Tyramide Signal Amplification systems (PerkinElmer #: NEL75300 1KT). Slides were visualized on a Zeiss LSM 700 confocal microscope. Images were acquired with identical parameters were used to compare sections from different genotypes.
Superscript 2 reverse transcriptase first strand cdna synthesis kit
Manufactured by Thermo Fisher Scientific
Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit is a reagent kit used for the synthesis of first-strand cDNA from RNA templates. The kit contains the Superscript II reverse transcriptase enzyme, buffer, and other necessary components for the conversion of RNA to cDNA.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using superscript 2 reverse transcriptase first strand cdna synthesis kit
1
In Situ Hybridization of Neuronal Markers
2
Transcriptional Profiling of Math5 in Retina and dLGN
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Retina and dLGN tissue were harvested from C57/BL6 mice at different embryonic and postnatal ages using methods described elsewhere [51 (link)]. RNA was isolated using the Bio-Rad Total RNA Extraction from Fibrous and Fatty Tissue kit (Bio-Rad). Reverse transcription and cDNA generation were made using Superscript II Reverse Transcriptase First- Strand cDNA Synthesis kit (Invitrogen). The following math5 primer pairs were used: 5′- ATGGCGCTCAGCTACATCAT- 3′ and 5′-GGGTCTACCTGGAGCCTAGC- 3′.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!