ABI PRISM® SNaPshot™ Multiplex Kit Quick Reference Card
Contents
1. LIZ size standard into each tube 3 Vortex briefly and quick spin 4 Denature the samples by placing them at 95 C for 5 minutes 5 Quick spin or tap the tubes or plates to bring liquid to the bottom of the tubes 6 Place the samples at 4 C until ready to load 7 To run samples on the 310 Genetic Analyzer verify that you have selected the following in the data collection software a GS STR POP 4 1 mL E5 module b E5 matrix c GeneScan 120 LIZ size standard change the default color to orange To run samples on the 3100 Genetic Analyzer verify that you have selected the following in the data collection software a E5 dye set b SNP36_POP4 module c GeneScan 120 LIZ size standard Analysis change the default color to orange To run samples on the 3700 DNA Analyzer verify that you have selected the following in the data collection software a E5 dye set b Modified SNP1_1 POP5 module c GeneScan 120 LIZ size standard Analysis change the default color to orange Kit Number of Reactions Part Number ABI Prism SNaPshot Multiplex Kit 1003 4323151 10002 4323154 50008 4323155 a Contains Multiplex Control Template and Multiplex Control Primer Mix for 30 control reactions Note The fluorescent dyes are assigned to the individual ddNTPs as follows ddNTP Dye Label Color of Analyzed Data A dR6G Green Cc dTAMRA Black G dR110 Blue T U dROX Red P N 4323975 Rev A AS Applied Bio2. ABI PRISM SNaPshot Multiplex Kit Quick Reference Card This quick reference guide is intended for use only by appropriately trained laboratory personnel who have read the user s manual and are experienced with the protocol For safety guidelines please refer to the ABI PRISM SNaPshot Multiplex Kit Protocol P N 4323357 For all chemcials in bold type below please read the MSDS and follow the handling instructions Wear appropriate protective eyewear clothing and gloves Preparing the SNaPshot Control Reactions Step Action 1 Label two 0 2 mL MicroAmp tubes One for the positive control reaction One for the negative control reaction 2 Combine the following items in each tube Positive Negative Control Control Item uL HL SNaPshot Multiplex Ready Reaction Mix 5 5 SNaPshot Multiplex Control Primer Mix 1 1 SNaPshot Multiplex Control Template 2 0 Deionized water 2 4 Total 10 10 3 Mix well spin briefly and immediately place on ice Preparing the SNaPshot Reactions Step Action 1 Combine the following ingredients on ice One Sample Item HL SNaPshot Multiplex Ready Reaction Mix 5 Pooled PCR products 3 Pooled SNaPshot primers 1 Deionized water 1 Total 10 2 Aliquot 10 uL into each MicroAmp tube well and immediately place on ice Thermal Cycling the SNaPshot Reactions Step Action 1 P
3. lace the tubes in a GeneAmp 9600 thermal cycler and set the volume to 10 yL 2 Repeat the following for 25 cycles 96 C for 10 seconds 50 C for 5 seconds 60 C for 30 seconds Note The conditions can be modified to accommodate specific primers 3 Rapid thermal ramp to 4 C and hold until ready for post extension treatment 4 Continue with Post Extension Treatment on the back of this card AS Applied Biosystems Post Extension Treatment Step Action 1 Add one of the following to the reaction mixture and incubate at 37 C for 1 hour 1 0 Unit of Shrimp AlkalinePhosphatase SAP or 1 0 Unit of Calf Intestinal Phosphatase CIP 2 Deactivate the enzyme by incubating at 75 C for 15 minutes 3 Place the sample at 4 C Preparing the SNaPshot Products for the ABI PRISM 310 Genetic Analyzer 3100 Genetic Analyzer or 3700 DNA Analyzer Step Action 1 Add 9 uL of HI Di formamide into each tube AEAN CHEMICAL HAZARD Formamide is harmful if absorbed through the skin and may cause irritation to the eyes skin and respiratory tract It may cause damage to the central nervous system and the male and female reproductive systems and is a possible birth defect hazard Please read the MSDS and follow the handling instructions Wear appropriate protective eyewear clothing and gloves 2 Add 0 5 uL of SNaPshot products and 0 5 uL of GeneScan 120
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