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Site Directed Mutagenesis (SDM) Rat

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Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Rat - Brain

Products Sigma-Aldrich Imprint® Chromatin Immunoprecipitation Kit

Get tips on using Human SCF ELISA Kit (ab100636) to perform ELISA Rat - SC

Products Abcam Human SCF ELISA Kit (ab100636)

Get tips on using HSP70 High Sensitivity ELISA kit to perform ELISA Rat - HSP70

Products Enzo Life Sciences HSP70 High Sensitivity ELISA kit

Get tips on using Total BDNF Quantikine ELISA Kit to perform ELISA Rat - BDNF

Products R&D Systems Total BDNF Quantikine ELISA Kit

Get tips on using Silencer®_Faslg siRNA (r) to perform siRNA / miRNA gene silencing Rat - F98 Faslg

Products Thermo Fisher Scientific Silencer®_Faslg siRNA (r)

Get tips on using Silencer® Select_Alkbh1 siRNA (r) to perform siRNA / miRNA gene silencing Rat - B35 Alkbh1

Products Thermo Fisher Scientific Silencer® Select_Alkbh1 siRNA (r)

Get tips on using Gibco™ DMEM, high glucose to perform Stem cell culture media Rat TSPCs

Products Fisher Scientific Gibco™ DMEM, high glucose

Get tips on using Gibco™ DMEM, high glucose to perform Stem cell culture media Rat BMSC

Products Fisher Scientific Gibco™ DMEM, high glucose

A restriction enzyme or restriction endonuclease is defined as a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at or near that site, known as restriction site or target sequence. The four most common types of restriction enzymes include: Type I (cleaves at sites remote from a recognition site), Type II (cleaves within or at short specific distances from a recognition site), Type III (cleave at sites a short distance from a recognition site), and Type IV (targets modified DNA- methylated, hydroxymethylated and glucosyl-hydroxymethylated DNA). The most common challenges with restriction digest include- 1. inactivation of the enzyme, 2. incomplete or no digestion, and 3. unexpected cleavage. The enzyme should always be stored at -20C and multiple freeze-thaw cycles should be avoided in order to maintain optimal activity. Always use a control DNA digestion with the enzyme to ensure adequate activity (to avoid interference due to high glycerol in the enzyme). For complete digestion, make sure that the enzyme volume is 1/10th of the total reaction volume, the optimal temperature is constantly maintained throughout the reaction, the total reaction time is appropriately calculated based on the amount of DNA to be digested, appropriate buffers should be used to ensure maximal enzymatic activity, and in case of a double digest, make sure that the two restriction sites are far enough so that the activity of one enzyme cannot interfere with the activity of the other. Star activity (or off-target cleavage) and incomplete cleavage are potential challenges which may occur due to suboptimal enzymatic conditions or inappropriate enzyme storage. To avoid these, follow the recommended guidelines for storage and reactions, and always check for the efficacy of digestion along with purification of digested products on an agarose gel.

Proteins Restriction Enzymes HindIII

A restriction enzyme or restriction endonuclease is defined as a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at or near that site, known as restriction site or target sequence. The four most common types of restriction enzymes inclue: Type I (cleaves at sites remote from a recognition site), Type II (cleaves within or at short specific distances from a recognition site), Type III (cleave at sites a short distance from a recognition site), and Type IV (targets modified DNA- methylated, hydroxymethylated and glucosyl-hydroxymethylated DNA). The most common challenges with restriction digest include- 1. inactivation of enzyme, 2. incomplete or no digestion, and 3. unexpected cleavage. The enzyme should always be stored at -20C and multiple freeze-thaw cycles should be avoided in order to maintain optimal activity. Always use a control DNA digestion with the enzyme to ensure adequate activity (to avoid interference due to high glycerol in the enzyme). For complete digestion, make sure that the enzyme volume is 1/10th of the total reaction volume, optimal temperature is constantly maintained throughout the reaction, the total reaction time is appropriately calculated based on the amount of DNA to be digested, appropriate buffers should be used to ensure maximal enzymatic activity, and in case of a double digest, make sure that the two restriction sites are far enough so that the activity of one enzyme cannot interfere with activity of the other. Star activity (or off-target cleavage) and incomplete cleavage are potential challenges which may occur due to suboptimal enzymatic conditions or inappropriate enzyme storage. To avoid these, follow the recommended guidelines for storage and reactions, and always check for the efficacy of digestion along with purification of digested products on an agarose gel.

Proteins Restriction Enzymes BamHI

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